Zebrafish has unique advantages over other animal models in the aspect of drug screens for active ingredients and gains more and more attentions in drug research and development recently. Thus, this article reviews the recent advance of zebrafish-based drug screens in Chinese traditional medicine (TCM) effective part research, monomer drug screening, activity evaluation of natural products, discovery of new uses for old drugs, and toxicity assessment in early-phase drug discovery.
Animals ; Drug Evaluation, Preclinical ; instrumentation ; methods ; Drugs, Chinese Herbal ; pharmacology ; Models, Animal ; Zebrafish ; growth & development

Surface plasmon resonance (SPR) biosensors have become an advanced method for measuring biological molecular interaction. This article is focused on the principle and advantages of SPR biosensor chip technology, and its new applications in biomedical research fields. Its application prospects are also discussed.
Clinical Laboratory Techniques ; Drug Evaluation, Preclinical ; methods ; Surface Plasmon Resonance ; instrumentation

We developed a novel microfluidic cell chip, which enabled drug delivery, fluid control and cell co-culture. The device consisted of an array of 6x6 cell culture chambers, a drug gradient generator and fluidic control valves. Micro-dam structures of the chambers were able to trap cells while loading and drug gradient network generated drug gradient of 6 different concentrations. Also we applied hydraulic valves to control the microfluid and simulate the microenvironment of cells. We had investigated the viability of co-culturing cells in the chip and the ability for drug screening. This microfluidic cell chip has the potential in cell-based research of high throughput drug screening.
Biosensing Techniques ; instrumentation ; methods ; Cells, Cultured ; Drug Evaluation, Preclinical ; methods ; Endothelial Cells ; cytology ; Hepatocytes ; cytology ; Humans ; Microfluidic Analytical Techniques ; instrumentation ; methods ; Microfluidics ; instrumentation ; methods ; Umbilical Veins ; cytology

OBJECTIVETo establish an in vitro screening model for steroid 5 alpha-reductase inhibitors using the microplate reader.

METHODSSteroid 5 alpha-reductase was obtained from the liver of female rats, an in vitro screening model for steroid 5 alpha-reductase inhibitors established using the 96-well plate and microplate reader after determination of the enzymatic activity, and the reliability of the model verified with the known 5 alpha-reductase inhibitors epristeride and finasteride. Added to the 96-well plate were the final concentrations of testosterone (0-40 micromol/L), NADPH (22 micromol/L), epristeride (0-60 nmol/L) or finasteride (0-60 nmol/ L) and steroid 5 alpha-reductase (20 microl), the total volume of each well adjusted to 200 microl with Tris-Hcl buffer. The 96-well plate was placed in the microplate reader, mixed and incubated at 37 degrees C, followed by detection of the A340nm value at 0 and 10 min and analysis of the data.

RESULTSThe Km value of steroid 5 alpha-reductase was 3.794 micromol/L, with a Vmax of 0.271 micromol/(L. min). The Ki of epristeride was 148.2 nmol/L, with an IC50 of 31.5 nmol/L, and the enzymatic reaction kinetic curve suggested that epristeride was an uncompetitive enzyme inhibitor. The Ki of finasteride was 158. 8 nmol/L, with an IC50 of 13.6 nmol/L. The enzymatic reaction kinetic curve showed that both epristeride and finasteride were competitive enzyme inhibitors, similar to those reported in the published literature.

CONCLUSIONA screening model was successfully established, which could rapidly and effectively screen steroid 5 alpha-reductase inhibitors in vitro.

5-alpha Reductase Inhibitors ; analysis ; Animals ; Drug Evaluation, Preclinical ; methods ; Female ; High-Throughput Screening Assays ; instrumentation ; methods ; Immunoenzyme Techniques ; Rats ; Rats, Sprague-Dawley

Leishmaniasis is a worldwide uncontrolled parasitic disease due to the lack of effective drug and vaccine. To speed up effective drug development, we need powerful methods to rapidly assess drug effectiveness against the intracellular form of Leishmania in high throughput assays. Reporter gene technology has proven to be an excellent tool for drug screening in vitro. The effects of reporter proteins on parasite infectivity should be identified both in vitro and in vivo. In this research, we initially compared the infectivity rate of recombinant Leishmania major expressing stably enhanced green fluorescent protein (EGFP) alone or EGFP-luciferase (EGFP-LUC) with the wild-type strain. Next, we evaluated the sensitivity of these parasites to amphotericin B (AmB) as a standard drug in 2 parasitic phases, promastigote and amastigote. This comparison was made by MTT and nitric oxide (NO) assay and by quantifying the specific signals derived from reporter genes like EGFP intensity and luciferase activity. To study the amastigote form, both B10R and THP-1 macrophage cell lines were infected in the stationary phase and were exposed to AmB at different time points. Our results clearly revealed that the 3 parasite lines had similar in vitro infectivity rates with comparable parasite-induced levels of NO following interferon-gamma/lipopolysaccharide induction. Based on our results we proposed the more reporter gene, the faster and more sensitive evaluation of the drug efficiency.
Amphotericin B/*pharmacology ; Animals ; Antiprotozoal Agents/*pharmacology ; Drug Evaluation, Preclinical/instrumentation/*methods ; Female ; Gene Expression ; Genes, Reporter ; Green Fluorescent Proteins/genetics/*metabolism ; Humans ; Leishmania major/*drug effects/genetics/growth & development/physiology ; Leishmaniasis, Cutaneous/*parasitology ; Luciferases/genetics/*metabolism ; Mice

OBJECTIVETo probe into the feasibility of screening anti-HIV compounds by using HIV-1 p24 detection kit made by Hebei Medical University.

METHODSThe sensitivity, reproducibility and efficacy of the Hebei p24 kit were evaluated compared with the commercially available Vironostika HIV-1 Antigen Microelisa System (Biomerieux).

RESULTSHebei p24 kit had high sensitivity and good reproducibility. In vitro screening demonstrated that there was no statistically significant difference (P greater than 0.05) between these two kits in assessing anti-HIV compounds.

CONCLUSIONHebei p24 kit could be used as an easily affordable alternative method for detection of HIV-1 in screening anti-HIV compounds.

Anti-HIV Agents ; isolation & purification ; pharmacology ; Cell Line ; Drug Evaluation, Preclinical ; instrumentation ; methods ; Feasibility Studies ; HIV Core Protein p24 ; analysis ; HIV-1 ; drug effects ; growth & development ; immunology ; Humans ; Reagent Kits, Diagnostic ; standards ; Reproducibility of Results