Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy / 中华眼底病杂志

Objective To investigate the inhibitory effect oflentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygeninduced retinopathy (OIR).Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group,simple OIR model group,OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group),with 16,32,32 and 32 mice,respectively.When the mice were 7 days old,the mice in the normal control group were fed in a routine environment,and the mice in the OIR model group,Vec group and PSF group were established OIR model.The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1 × 10~ TU/ml) at the age of 12 days.No injection was performed in the normal control group and simple OIR group.RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina.Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase1 (HO-1).Western blot analysis was applied to detect the protein expression ofNrf2,HO-1 and PSF.Results Of the normal control group,simple OIR model group,Vec group and PSF group,the number of pre-retinal neovascular cell nuclei were 0.00,14.36 ± 5.50,15.67 ± 4.96,8.13 ± 2.09,the non-perfusion area were 0.00％,(35.71 ± 2.81)％,(36.57 ± 4.53)％,(15.33 ± 4.75)％,respectively.The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87,165.70;P＜0.05).Compared with the normal control group,there were more pre-retinal neovascular cell nucleis and larger nonperfusion area in the simple OIR model group and Vec group (P＜0.05).Compared with the simple OIR model group and Vec group,there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05).Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2,HO-1 (F=53.66,83.54) and protein expression ofNrf2,HO-1 and PSF (F=58.38,52.69,24.79) among 4 groups were significant (P＜0.05).The rnRNA expression ofNrf2,HO-1 and protein expression of Nrf2,HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05).The mRNA expression ofNrf2,HO-1 and protein expression ofNrf2,HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05).model group and Vec group (P＜0.05).Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.

Interaction between human serum albumin and cholesterol-grafted polyglutamate as the potential carriers of protein drugs

Currently there is no successful platform technology for the sustained release of protein drugs. It seems inevitable to specifically develop new materials for such purpose, and hence the understanding of protein-material interactions is highly desirable. In this study, we synthesized cholesterol-grafted polyglutamate (PGA--Chol) as a hydrophobically-modified polypeptide, and thoroughly characterized its interaction with a model protein (human serum albumin) in the aqueous solution by using circular dichroism, fluorescence methods, and light scattering. With the protein concentration fixed at 5 μmol/L, adding PGA--Chol polymers into the solution resulted in continuous blue shift of the protein fluorescence (from 339 to 332 nm), until the polymer molar concentration reached the same value as the protein. In contrast, the un-modified polyglutamate polymers apparently neither affected the protein microenvironment nor formed aggregates. Based on the experimental data, we proposed a physical picture for such protein-polymer systems, where the polymer first bind with the protein in a 1:1 molar ratio a fraction of their hydrophobic pendant cholesterol resides along the polymer chain. In this protein/polymer complex, there are excess unbound cholesterol residues. As the polymer concentration increases, the polymers form multi-polymer aggregates around 200 nm in diameter the same hydrophobic cholesterol residues. The protein/polymer complex also participate in the aggregation their excess cholesterol residues, and consequently the proteins are encapsulated into the nanoparticles. The encapsulation was also found to increase the thermal stability of the model protein.

Development strategies and application of Fc fusion protein / 中华微生物学和免疫学杂志

Monoclonal antibodies ( McAb ) , a rapidly expanding class of therapeutic biological products, have been widely used in the treatment of various diseases. Although antibodies have shown tre-mendous clinical value, the breadth and complexity of human diseases urge researchers to constantly develop new drugs with new modes of action rather than to confine them to monoclonal antibodies alone. Fc fusion proteins are becoming a new research hotspot and have shown great potential in the treatment of many human diseases. At present, Fc fusion proteins on the market usually take the interaction between receptors and lig-ands as the main guiding strategy to develop more safer and effective clinical drugs. This article mainly fo-cused on the ligand-receptor interaction as the drug design strategy and the application. of Fc fusion pro-teins.

Advances in the Research of Long Life Recombinant Protein Drug / 中国生物工程杂志

The recombinant protein drug usually has a short half-life after intravenous (IV) or subcutaneous (SC) administration. The methods of prolonging the half-life of recombinant protein drug in common use are mainly based on three principles: 1 Amplifying the molecule weight of protein drug; 2 Making use of drug balance in the blood; 3 Reducing Immunogenicity. Three methods were focused on: Analog construction, PEGylation and Albumin fusion technology. The characteristics, half-life and immunogenicity problem of their products in the market and under development are summarized.

"Reverse Immunology" and The Discovery of Immune-related Functional Genes / 生物化学与生物物理进展

The completion of sequencing human genome creates a new era of biological science and technology. Although the sequence of the human genome has been known, it is still hard to rapidly explore the whole functional genes, especially, their interaction with each other and the meaning to the body. However, the "reverse biology" which comes into being in the recent years provides us a series of novel ideas and technologies for discovering new functional gene, among which the immune-related genes have attracted more attentions, clarifying how functional gene works and their potential value in application.

Study on Self-Assembling Peptide In-Situ Gel as Potential Protein Carrier / 中国药房

OBJECTIVE: To investigate the feasibility of introducing self-assembling peptides as protein drug carrier.METHODS: By using RAD16-Ⅰ as a model of self-assembling peptides,lysozyme as the model of protein drug,the property of self-assembling peptide solution containing protein to form hydrogel in-situ were characterized by rheology,and the property of protein release in vitro from self-assembling peptide in-situ hydrogel was studied.RESULTS: The self-assembling peptide RAD16-Ⅰ can form hydrogel with certain mechanical intensity rapidly after its solution contained protein being mixed with PBS.Proteins can release easily from RAD16-Ⅰ hydrogel in a sustained-release manner to some extent.About 80% of proteins can be released from the hydrogel within 8 hours and the cumulative protein release from 1.5% hydrogel was less than 90%.The bio-activity of lysozyme released from RAD16-Ⅰ hydrogel was well preserved as 98% to 115%.CONCLUSION: Self-assembling peptide can be employed as potential in-situ hydrogel carrier for protein drugs.

Effects of Qixue Bingzhi Prescription on Hypersensitive C-reactive Protein and Other Indexes in the Patient of Cervical Atherosclerosis / 中医杂志

Objective:To study intervenient action of Qixue Bingzhi Prescription on the patient of cervical atherosclerosis(AS). Methods:63 cases of cervical AS were randomly divided into a treatment group(n=33) and a control group(n=30), with 25 cases of same age ranges used as healthy group. The control group was treated with basic treatment of western medicine and the treatment group with the basic treatment plus Qixue Bingzhi Prescription for one month. Changes of blood lipid metabolism, interleukin levels, supersensitive C-reactive protein(Hs-CRP) content and the leukocyte adherence expression rate were observed. Results: The total cholesterol (TC), triglyceride(TG), AS index (TC/HDL-C), interleukin-6 (IL-6), interleukin-8 (IL-8) and Hs-CRP levels and lymphocytes and the neutrophilic granulocyte adhesion molecule expression rate (ICD11b and zCD11b) decreased significantly(P 0. 05), Conclusion: Qixue Bingzhi Prescription can effectively and safely intervene in blood lipid metabolism and inflammatory reaction in the patient of cervical AS, and it has prospect of development in preventing formation of AS and stabilizing plaque.