Phage peptide libraries are collections of the specific length of short peptides,and they are based on phages as their carriers.They include mimotope libraries and peptide antibody libraries.Phage-displayed peptide libraries have been used to isolate specific ligands for numerous protein targets,and they have been proven useful in defining antigen momitopes,rapid determination of binding energetics at protein-protein interfaces,designing of vaccine and tumor research aspects.This review summarized the research progression of phage peptide library.

Pyrrolizidine alkaloids (PAs) are widely distributed in many plants including medicinal herbs. The hepatotoxicity of PAs has been known academically for a long time, however, their reproductive toxicity, mutagenesis and carcinogenicity have been less researched. This article is an overview of the clinical and experimental reports of the reproductive toxicity, mutagenesis and carcinogenicity of PAs, the effective factors and generating mechanism of the toxicity.
Animals ; Biomedical Research ; Humans ; Plant Extracts ; analysis ; toxicity ; Plants, Medicinal ; chemistry ; toxicity ; Pyrrolizidine Alkaloids ; analysis ; toxicity

AIM and METHODS:The Ph.D.-7 phage display library was used to isolate peptides specific for glioma SWO-38 cell by whole cell screening.Moreover,binding efficiency analysis was carried out to test the binding specificity of the clones obtained. RESULTS: After three rounds of biopanning,a high concentration of phage clones was obtained and two of them were found to be highly specific to glioma SWO-38. CONCLUSION: Highly specific clones against neurtral glioma cells can be obtained from a phage display library by simple procedures.

OBJECTIVETo investigate the fetotoxicity of monocrotaline.

METHODMouse whole embryo culture (WEC) was applied. Post-implantation (8.5 d) mouse embryos were isolated from their mothers and put into the medium of immediately centrifuged serum (ICS) prepared from rats. Different concentrations of monocrotaline (100, 50, 25, 12.5 mg x L(-1)) were added into the WEC. Development (yolk sac diameter, crown-rump length, head length, somite number) and organic morphodifferentiation (yolk sac circulation, allantois, embryonic flexion, heart, brain, optic-otic-olfactory organ, branchial arch, maxillary, mandible, bud) of embryos were observed at 48 h after treatment.

RESULTObvious fetotoxicity could be observed in various monocrotaline treatment groups in a dose-dependent manner. Development of embryos was delayed significantly at dose 12.5-100 mg x L(-1). Malformations were shown in all organic morphodifferentiation indice, especially in opti-otic organ, mandible and bud.

CONCLUSIONMonocrotaline had obvious fetotoxicity in vitro WEC, indicating that exposure of pregnant mice to monocrotaline may have potential risk on fetus.

Animals ; Cell Differentiation ; drug effects ; Culture Media ; Embryo, Mammalian ; drug effects ; physiology ; Female ; Male ; Mice ; Monocrotaline ; toxicity

OBJECTIVEBy using RAW 264.7 macrophage cell line, we studied the dose-effect relationship of endotoxin induced RAW 264.7 cells to release TNF-alpha, and then detected the content of endotoxin in 8 kinds of injections, so that we can investigate the feasibility and the interference factors of the novel test.

METHODBy using endotoxin of different concentrations to induce RAW 264. 7 cells to release TNF-a, we drew the curve of dose-effect relationship between endotoxin and generated TNF-alpha. Then we detected the content of TNF-alpha in yuxingcao, shuanghuanglian, qingkailing, gegensu, xiangdan, qianrongmei and jiangxianmei injections and shuanghuanglian powder injection, and calculated their content of endotoxin.

RESULTThe endotoxin could induce the cells to release TNF-alpha in a good dose-dependent manner, even at a very low concentration. In the range of maximum available dilution multiple, the content of endotoxin in the rest 7 kinds of injections was less than 1.0 EU x mL(-1) except qingkailing injection of two batch.

CONCLUSIONCytokine revulsion has the advantage of wide detection range, high sensitivity, simple operation, and the detected endotoxin is of bioactivity. This method provides another technical mean for pyrogen test of injections.

Animals ; Biological Assay ; methods ; Cell Line ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Endotoxins ; adverse effects ; analysis ; Macrophages ; drug effects ; immunology ; Mice ; Tumor Necrosis Factor-alpha ; analysis ; immunology

OBJECTIVETo detect content of bacterial endotoxin in Yuxingcao and Qingkailing injections by specific and nonspecific tachypleus amebocyte lysate technique for in order to investigate the feasibility of specific tachypleus amebocyte lysate technique for detecting bacterial endotoxin in traditional Chinese drug injections.

METHODDifferent batches of Yuxingcao and Qingkailing injections were detected by specific and nonspecific tachypleus amebocyte lysate kits.

RESULTYuxingcao injection could be detected by specific and nonspecific tachypleus amebocyte lysate technique, Whereas Qingkailing injection could be detected only by specific tachypleus amebocyte lysate.

CONCLUSIONUsing specific tachypleus amebocyte lysate as a substitute for nonspecific tachypleus amebocyte lysate is an effective method for detecting content of bacterial endotoxin in Qingkailing injection.

Animals ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Endotoxins ; analysis ; Horseshoe Crabs ; Limulus Test ; methods

OBJECTIVETo establish a model of gastric precancerous lesion by using Aristolochic manshuriensis which contains aristolochic acids.

METHODThe SD rats were randomly divided into four groups: control and three different doses of ethanol extractive of A. manshuriensis (EEA) (corresponding to aristolochic acid I 2.5, 5.0, 10.0 mg x kg(-1)), respectively. EEA was intragastrically given to rats every other day. At the end of the 10th, 15th, 20th week, part of the rats in each group was sacrificed and the stomachs were weighed. The gastric tumor was assessed by the weight and the relative stomach weight to the body weight. The stomachs were fixed in 4% neutral formalin, and the paraffin imbedding tissues were sliced and HE stained. Histomorphology was observed under the light microscope to determine gastric hyperplasia, mucosa precancerosis (atypical hyperplasia) and gastric cancer formation.

RESULTThe rats treated with different doses of EEA for 10 weeks induced mucosa papillary, epithelioma hyperplasia. Histological observation showed mucosa precancerosis lesions characterized as atypical hyperplasia at the dose levels corresponding to aristolochic acid I 5.0 and 10.0 mg x kg(-1) treated for 10 weeks. The incidence rate of gastric precancerosis in those two groups was 100% at the 15th week. Malignant tumors were observed in most of the animals in 10.0 mg x kg(-1) group. The animals in 5.0 mg x kg(-1) group were well tolerant compared to 10.0 mg x kg(-1) group during the course of experiment, so the dose of aristolochic acid I 5.0 mg x kg(-1) and 10-15 weeks treatment were considered to be optimum to establish the model of gastric precancerosis.

CONCLUSIONA rat model of gastric precancerosis can be induced within a short duration by giving an oral administration of the ethanol extract of A. manshuriensis which contains aristolochic acids.

Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; administration & dosage ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Stomach Neoplasms ; drug therapy ; pathology

OBJECTIVETo investigate the antiatherogenic effect and possible mechanisms of the extracts of Radix Salviae Miltiorrhizae (RSM) or Fructus Crataegi (FC), as well as their interaction.

METHODWistar rats were randomly divided into 2 groups: normal group and model group. The atherosclerotic model rats were injected VD3 and ovalbumin, while fed with high cholesterol diet. After the model was determined successfully, all model rats were divided into normal group, model group, Xuezhikang group, RSM group, FC group, mixture of RSM and FC group. Each group was given the corresponding drugs for 4 weeks. After 12 weeks, blood serum were analyzed for total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C), superoxide dismutase ( SOD), malondialdehyde (MDA) and nitric oxide (NO). And the blood plasma also analyzed for levels of endothelin (ET), 6-keto prostaglandin F1alpha (6-keto-PGF1alpha), thromboxane B2 (TXB2), C-reactive protein (CRP), interleukin 6 (IL-6), interleukin 8 (IL-8), tumor necrosis factor alpha (TNF-alpha) and so on. At last, the pathological observation of aorta was carried out.

RESULTCompared with those in model group, the TC, TG, LDL-C, ET, TXB2 and MDA levels and TXB2/PGF1alpha ratio were reduced, while the HDL-C, the serum SOD, No and 6-keto-PGF1alpha level were raised in the intervention groups. Although the levels of CRP, IL-6 and IL-8 were lower than model group, there was no obvious effect on the releasing of TNF-alpha.

CONCLUSIONRSM and FC could inhibit the atherogenesis formation and development, which might be due to regulating the lipid metabolism, enhancing the antioxidation, and reducing the release of inflammatory factors.

Animals ; Atherosclerosis ; prevention & control ; C-Reactive Protein ; analysis ; Crataegus ; Disease Models, Animal ; Interleukin-6 ; blood ; Lipid Peroxidation ; drug effects ; Lipids ; blood ; Male ; Plant Extracts ; therapeutic use ; Rats ; Rats, Wistar ; Salvia miltiorrhiza

OBJECTIVETo modify the empirical method of precision-cut liver slice technique, and study the hepatotoxicity of monocrotaline by this technique.

METHODLiver slices were prepared by the domestic shaking slicer. The technique of precision-cut liver slice was established by detecting MTT reduction used as the slice viability under different culture medium, thickness of slices, pH and culture temperature. After monocrotaline and liver slices co-culture for 6, 24 h, the slice viability, enzyme activity of GPT, GOT, LDH, GGT and protein concentration were detected by MTT reduction, enzyme kinetics method and BCA protein assay method, respectively.

RESULTWhen the thickness of slices was 200 microm and pH of medium was 6.8, culture temperature was 37 degrees C, BPM culture medium, the viability of slices could maintain on a steady level. LDH leakage was significantly increased and protein content was obviously decreased after monocrotaline co-culture for 24 h with final concentration 0.02, 0.1 and 0.5 g x L(-1). No statistically significant difference between control group and monocrotaline 3 dose groups was observed in the slice viability and the content of GPT, GOT, LDH, GGT and protein after monocrotaline co-culture for 6 h.

CONCLUSIONThe slice viability could retain 24 h in modified BPM medium surroundings; monocrotaline displayed liver toxicity in some degree after co-culture for 24 hours in 0.02, 0.1 and 0.5 g x L(-1) concentration.

Animals ; Hydrogen-Ion Concentration ; L-Lactate Dehydrogenase ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Monocrotaline ; toxicity ; Rats ; Rats, Sprague-Dawley ; Temperature ; Toxicity Tests

OBJECTIVETo establish a simple and feasible method of anaphylactoid test on awaked small animals for screening and assessing anaphylactoid reaction of traditional Chinese medicine (TCM) injection with different concentration of tween 80.

METHODTest substances containing 0.4% Evans blue were intravenously injected into mice at volume of 20 mL x kg(-1) or guinea pigs at a volume of 30 mL x kg(-1). The behaviors were observed and the vascular permeability of ears evaluated by the extent of ear blue staining and absorbance of Evans blue extraction of ears were tested at 30 min after injection.

RESULTTween 80 solution, Yuxingcao injection with tween 80, and Shuanghuanglian powder injection obviously increased vascular permeability of ears characterized as ear blue staining and increased absorbance of the Evans blue extract from ears extracted by acetone saline both in mice and in guinea pigs in a concentration-dependent (in the case of tween 80) or a dose-dependent (Shuanghuanglian) manner.

CONCLUSIONEar vascular permeability test in mice and guinea pigs can be used as animal models to screen and test anaphylactoid reaction induced by injections.

Anaphylaxis ; chemically induced ; Animals ; Capillary Permeability ; drug effects ; Guinea Pigs ; Male ; Medicine, Chinese Traditional ; adverse effects ; Mice ; Mice, Inbred ICR ; Models, Animal