Natural products are valuable resources for discovering new drugs. So far, screening bioactive compounds from organism extracts is still an important and challenging task. Traditional biometric guided method involves repeated fractionation steps and bioactivity tests, which are time-consuming, labor-consuming, and inefficient. Ligand fishing is a bioanalysis method for screening ligands from complex organism extracts based on intermolecular affinity interactions. It has the characteristics of strong specificity, high efficiency, and less requirement for sample pretreatment. In this review, we summarize the classification of ligand fishing strategy and its application in enzyme inhibitors screening. Finally, the development prospects of this technology are forecasted.

Objective: To observe the effects of needles with different diameters on the gastrointestinal function in mice. Methods: Eighteen Kunming mice were randomly divided into group of 0.25 mm needle, group of 0.35 mm needle, and control group. The acupoint of Zusanli (ST 36) was needled once a day for 5 days. The effects of needles with different diameters were observed by measuring the distance of the carbon moved in the intestine. Results: The distance of the carbon moved in the intestine was longer in the acupuncture group than in the control group (P<0.05), and it was longer in the group of 0.35 mm needle than in the group of 0.30 mm needle, there was no significant difference (P>0.05). Conclusion: Acupuncture treatment can enhance the peristalsis function of stomach and intestine in mice. The diameter of needle has no effect on the gastrointestinal function.

OBJECTIVETo establish the HPLC fingerprint of Radix Aconiti Kusnezoffii.

METHODThe chromatographic separation was performed on a Kromasil C18 (4.6 mm x 250 mm, 5 microm) eluted with a mobile phase of acetonitrile-ammonium acetate buffer (2. 5 per thousand acetic acid-ammonia pH 10.5) (60:40). The UV detection wavelength was set at 240 nm and the flow rate was set at 1.0 mL x min(-1).

RESULTThe RSD of precision and repeatability was less than 2%. Under the selected chromatographic conditions, good HPLC fingerprints of Radix Aconiti Kusnezoffii were obtained.

CONCLUSIONThe method was simple, accurate and repeatable. It can be used for the quality control of Radix Aconiti Kusnezoffii.

Aconitine ; analogs & derivatives ; analysis ; Aconitum ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

To study the suppression effect of light rare earth elements (RE) on proliferation of two cancer cell lines. Two cancer cell lines PAMC82 and K562 were used to examine their colony-forming ability in soft agar, microtubule structure, calmodulin levels and regulation of some gene expressions by Northern blot analysis with and without treatment by RE. The results showed that on soft agar culture the colony-forming ability of human gastric cancer cell line PAMC82 treated by RE chloride decreased and the PAMC82 cell microtubule abnormal structure became normal. The calmodulin (CaM) levels decreased in human leukemia cells (K562) treated with cerium chloride and neodymium chloride. The Northern blot analysis revealed marked up-regulation of p53, p16(MTS1), p21(WAF1) gene expressions in PAMC82 cells treated with lanthanum chloride and cerium chloride, as compared to control PAMC82 cells. The light rare earth elements studied have certain suppression effects on proliferation of cancer cells. This effect might be related to the decrease of calmodulin and up-regulation of some gene expressions in cancer cells.

OBJECTIVETo evaluate the effect of post-treatment PSA kinetics on the prognosis of prostate cancer (PCa).

METHODSWe retrospectively reviewed the clinical data of 114 cases of locally advanced PCa treated by maximal androgen blockade (MAB) combined with brachytherapy, and analyzed the association of the changes in PSA kinetics with the prognosis of the patients.

RESULTSThe median survival time of the patients was 81 (15 - 144) months, with 1-, 3- and 5-year survival rates of 91. 23%, 78.07% and 68.42% , respectively. Univariate analysis indicated that the baseline PSA level, PSA nadir, the time of PSA decreasing to nadir, PSA doubling time, and the extent of PSA declining were all predictive factors for the survival time of the PCa patients. Multivariate analysis demonstrated that PSA nadir, the time of PSA decreasing to nadir, and the extent of PSA declining were three independent prognostic factors, which prolonged the long-term survival of the patients by 1.7, 3.2 and 6.8 times, respectively.

CONCLUSIONFor locally advanced PCa treated by MAB combined with brachytherapy, PSA nadir <1 micro g/L, the time to nadir <3 months, and the extent of PSA declining >96% are independent prognostic factors.

Aged ; Aged, 80 and over ; Androgens ; administration & dosage ; therapeutic use ; Brachytherapy ; Humans ; Male ; Middle Aged ; Prognosis ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms ; metabolism ; therapy ; Retrospective Studies

OBJECTIVETo evaluate the effect and possibility of surgical ablation of the pulmonary vein orifices under direct vision with transballoon ultrasound ablation catheter for patients with permanent atrial fibrillation and rheumatic valve disease.

METHODS21 consecutive patients with rheumatic valve disease and permanent atrial fibrillation undergoing mitral valve replacement surgery were enrolled for this study from December 2002 to September 2003. All the cases were divided into 2 groups by whether or not receiving an additive pulmonary vein ablation procedure. The test group [6 male, 5 female, aged (51.55 +/- 7.83) years, atrial fibrillation duration (5.50 +/- 5.40) years, left atrial diameter (7.27 +/- 1.39) cm, LVEF (53.95 +/- 4.54)% and NYHA class II - IV] undertook a surgical isolation of the pulmonary vein orifices by using a transballoon ultrasound ablation catheter addition to routine mitral valve replacement. The control group [3 male, 7 female, aged (53.30 +/- 7.86) years, atrial fibrillation duration (4.50 +/- 3.47) years, left atrial diameter (6.74 +/- 0.62) cm, LVEF (56.91 +/- 3.78)% and NYHA class II - IV] received the valve replacement surgery alone.

RESULTSThere were not any complications in both groups. With an electrical cardioversion 3 months after the surgery, 73% patients in the ultrasound ablation group were free from AF over 1 year while only 10% patients in control group (P=0.003). During an average follow-up duration of (45.92 +/- 4.61) months, 63.6% were in sinus rhythm in ultrasound ablation group while none in the control group. Left atrial volume decreased significantly at 1 year after surgery compared to that at 3 months after surgery in the test group [(97.83 +/- 32.39) cm(3) vs. (150.78 +/- 52.32) cm(3), P<0.05], and the end systolic diameter (LAESD) and end diastolic diameter (LAEDD) also decreased [(4.12 +/- 0.39) cm vs. (5.09 +/- 0.98) cm, P<0.05, respectively], while there were no apparently changes in the control group.

CONCLUSIONSAblation of the orifices of the pulmonary veins under direct vision with transballoon ultrasound ablation catheter during mitral valve surgery seems effective to maintain sinus rhythm after electrical cardioversion and could be performed safely. The function of left atrial and cardiac output improved during long term follow-up of 46 months.

Atrial Fibrillation ; therapy ; Catheter Ablation ; methods ; Catheterization ; Female ; Heart Valve Diseases ; therapy ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; Rheumatic Heart Disease ; therapy ; Ultrasonic Therapy

OBJECTIVETo explore Effects of marine collagen peptides (MCPs) on markers of metablic nuclear receptors, i.e peroxisome proliferator-activated receptor (PPARs), liver X receptor (LXRs) and farnesoid X receptor (FXRs) in type 2 diabetic patients with/without hypertension. METHOD Study population consisted of 200 type 2 diabetic patients with/without hypertension and 50 healthy subjects, all of whom were randomly assigned to MCPs-treated diabetics (n = 50), placebo-treated diabetics (n = 50), MCPs-treated diabetics with hypertension (n=50), placebo-treated diabetics with hypertension (n = 50), and healthy controls (n = 50). MCPs or placebo (water-soluble starch) were given daily before breakfast and bedtime over three months. Levels of free fatty acid, cytochrome P450, leptin, resistin, adiponectin, bradykinin, NO, and Prostacyclin were determined before intervention, and 1.5 months, and 3 months after intervention. Hypoglycemia and the endpoint events during the study were recorded and compared among the study groups.

RESULTAt the end of the study period, MCPs-treated patients showed marked improvement compared with patients receiving placebo. The protection exerted by MCPs seemed more profound in diabetics than in diabetics with hypertension. In particular, after MCPs intervention, levels of free fatty acid, hs-CRP, resistin, Prostacyclin decreased significantly in diabetics and tended to decrease in diabetic and hypertensive patients whereas levels of cytochrome P450, leptin, NO tended to decrease in diabetics with/without hypertension. Meanwhile, levels of adiponectin and bradykinin rose markedly in diabetics following MCPs administration.

CONCLUSIONMCPs could offer protection against diabetes and hypertension by affecting levels of molecules involved in diabetic and hypertensive pathogenesis. Regulation on metabolic nuclear receptors by MCPs may be the possible underlying mechanism for its observed effects in the study. Further study into its action may shed light on development of new drugs based on bioactive peptides from marine sources.

Adipokines ; blood ; Aged ; Biomarkers ; blood ; Bradykinin ; blood ; C-Reactive Protein ; metabolism ; Collagen ; therapeutic use ; Cytochrome P-450 Enzyme System ; blood ; Diabetes Mellitus, Type 2 ; blood ; complications ; drug therapy ; Epoprostenol ; blood ; Fatty Acids, Nonesterified ; blood ; Female ; Humans ; Hypertension ; blood ; complications ; drug therapy ; Male ; Marine Biology ; Middle Aged ; Nitric Oxide ; blood ; Peptides ; therapeutic use ; Prospective Studies ; Receptors, Cytoplasmic and Nuclear ; metabolism

Pseudoallergic reactions of Qingkailing injection (QKLI) was assessed by vascular hyperpermeability which were indicated by ear blue staining in ICR mice after single intravenous injection of QKLI mixed with Evans blue (EB) and skin blue spot formation in SD rats after intradermal injection of QKLI and intravenous injection of EB. In addition, QKLI-induced histamine, VEGF, TNF-alpha release was measured after ICR mice received the single dosing of QKLI iv. The mild vascular hyperpermeability characterized by ear blue staining could be observed in mice after intravenous injection of QKLI and EB. Intracutaneous injection of 50 micro L of test solution containing QKLI (25,50 microL) in rat back skin caused obvious local vascular hyperpermeability at the injection sites so as to result the larger diameters of blue spots than that in negative control group (P <0. 01). QKLI induced a significant increase of VEGF and a slight elevation of histamine in mice after intravenous administration, while TNF-alpha showed no change after QKLI iv. The results in this study indicated that both intravenous injection and intracutanous injection of QKLI could induce vascular hyperpemeability so as to cause pseudoallergic reaction in mice and rats. QKLI-induced pseudoallergic reaction may be associated with the release of histamine and VEGF.
Animals ; Drug Hypersensitivity ; blood ; etiology ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Histamine ; blood ; Injections ; Male ; Mice ; Rats ; Skin ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood