Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

OBJECTIVETo investigate effects of electro-acupuncture (EA) on Du Meridian on the water channel aquaporin-4 (AQP-4) expression and hind limbs function recovery in experimental spinal cord injured rats.

METHODSOne hundred and fifty SD rats were divided into the normal group, the model group and the EA group, with 50 rats in each group, modified Allen method was used to establish spinal cord injury model in the model and EA group. Rats in the EA group was EA on Dazhui and Ming men acupoints after modelling successfully. At the 1st, 3rd, 7th, 14th and 21th day after injury, the hind limbs function of rats was evaluated by BBB scales respectively, and the AQP-4 expression in spinal cord tissue was determined by immunohistochemistry technique and quantity analysed with image analyzer.

RESULTSAt the 1st day after spinal cord injury, the AQP-4 expression was significantly increased in gray matter and white matter of spinal cord in the model group and the EA group, which reached the peak at the 3rd day, but showed significant difference between the two groups (P < 0.05), and the difference at the 7th, 14th and 21th day became more significant (P < 0.01).

CONCLUSIONEA on Du Meridian can down-regulate AQP-4 expression after spinal cord injury, inhibit spinal cord edema for alleviating secondary spinal cord lesion, so as to protect the residual normal spinal cord tissues and promote the rebuilding of nervous tissues.

Animals ; Electroacupuncture ; Extremities ; physiopathology ; Female ; Male ; Meridians ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; therapy

To combine fingerprint and drug release rate in vitro, in order to study in vitro release of complex components of Chuanping sustained tablets, compound traditional Chinese medicine preparation. A qualitative determination of the characteristic peaks of the compound preparations were conducted by the fingerprint. The results of the dissolution rate determination under different release conditions showed that the release of three index components (methamphetamine, pseudoephedrine and scopolamine) of Chuanping sustained tablets was less affected by gastrointestinal factors, with similarity factors being more than 80 with unknown component release curves of three major characteristic peaks in the fingerprint. The qualitative determination proved that multiple components of the compound traditional Chinese medicine preparation was dissolved in vitro at similar rates, realizing the balanced release of complex components of the compound traditional Chinese medicine preparation. This study layed a theoretical and experimental basis for quality evaluation for the compound traditional Chinese medicine preparation.
Drugs, Chinese Herbal ; chemistry ; Tablets

BACKGROUND: Preliminary studies have found that ligustrazine can effectively improve the levels of nitric oxide and prostaglandin E2, and alleviate the inflammatory response of osteoarthritis, but the related mechanism remains unclear. OBJECTIVE: To observe the effect of ligustrazine on the expression levels of type Ⅱ collagen fiber α 1, vascular endothelial growth factor (VEGF) mRNA and miR20b in the articular cartilage of knee osteoarthritis model, and to explore the mechanism of ligustrazine for early-stage knee osteoarthritis in rats. METHODS: Healthy male Sprague-Dawley rats were randomized into normal, model, high- and low-dose ligustrazine, and positive control groups. Rats in the latter four groups were used to establish the model of early-stage knee osteoarthritis by intra-articular injection of papain, and were then intragastrically given the administration of normal saline, 100 and 50 mg/kg ligustrazine, and 24 mg/kg celecoxib, respectively. The normal group was given the same volume of normal saline. The treatment in each group lasted for 6 weeks. Then, the rat cartilage was taken, and changes of cartilage tissues were assessed by Mankin scores. Expression levels of type Ⅱ collagen fiber α 1, VEGF mRNA and miR20b in the cartilage were detected by qRT-PCR. RESULTS AND CONCLUSION: The order of Mankin scores was as follows: normal group < high-dose ligustrazine group < positive control group < low-dose ligustrazine group < model group (P < 0.05). The mRNA expression levels of type Ⅱ collagen fiber α 1 and VEGF were the highest in the normal group, followed by the high-dose ligustrazine group, positive control group, low-dose ligustrazine group, and model group (P < 0.05). The expression level of miR20b was significantly up-regulated except the model group, and its order was follows: high-dose ligustrazine group < positive control group < low-dose ligustrazine group. Our results indicate that ligustrazine has a positive effect on early-stage knee osteoarthritis in rats, and the possible mechanisms by which ligustrazine promotes cartilage repair are to up-regulate miR20b expression and to inhibit VEGF mRNA expression.

Objective:To observe the effect of Shaoyaotang on the contents of cell adhesion molecule-1 （ICAM-1） and transforming growth factor-β₁ （TGF-β₁） in serum of large intestine damp-heat syndrome of ulcerative colitis （UC） in rats， and the gene and protein expressions of leukocyte differentiation antigen14 （CD14）， Fas-related death domain protein （FADD） and cysteinyl aspartate specific protease-8 （Caspase-8） in the focal colon tissue. Method:A total of 80 SPF Wistar rats were randomly divided into the blank group （n=10） and modeling group （n=70）. The large intestine damp-heat syndrome of UC rats was replicated by the combination of disease and syndrome， which was high-fat， high-sugar and spicy diets combined with 2， 4-dinitrobenzene sulfonic acid （DNBS） and ethanol. After successful modeling， the modeled groups were divided into model group， sulfasalazine （SASP）control group， and low， medium and high-dose Shaoyaotang groups by the method of random number table， with14 rats in each group. Low， medium and high doses of Sulfasalazine 0.2 g·kg^-1·d^-1 and Shaoyaotang （6， 12， 24 g·kg^-1·d^-1）were given by gavage. The blank group and the model group were given equal volume of normal saline for 21 days. The contents of serum ICAM-1 and TGF-β₁ were detected by enzyme-linked immunosorbent assay （ELISA）， the expressions of CD14， FADD and Caspase-8 mRNA in colon tissues were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）， and the expressions of CD14， FADD and Caspase-8 protein in colon tissues were detected by Western blot. Result:Compared with the blank group， the serum ICAM-1 level in the model group were significantly increased， whereas the content of TGF-β₁ were significantly decreased （P<0.05）. The relative expression levels of CD14， FADD， Caspase-8 mRNA and protein were significantly increased （P<0.05）. Compared with the model group， the content of ICAM-1 in the serum of the rats in the medium， high-dose Shaoyaotang groups and the SASP group were significantly decreased， while the content of TGF-β₁ in the serum of the rats in the low， medium， high-dose Shaoyaotang groups and the SASP group were significantly increased （P<0.05）. The expression levels of CD14， FADD， Caspase-8 mRNA and protein in each intervention group were significantly decreased （P<0.05）， especially in the high-dose Shaoyaotang group and the SASP group. Conclusion:Shaoyaotang has a certain intervention effect on UC rats with large intestine damp-heat syndrome， and its mechanism may be related to the inhibition of CD14， FADD and Caspase-8 genes and proteins expression.