Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.
Amino Acids ; analysis ; Animals ; Chickens ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Endothelium ; chemistry ; Gizzard, Avian ; chemistry

Objective To observe the effect of the 8-word bandage on the walking ability of stroke patients with knee hyperextension. Methods Fifty patients with stroke combined with knee hyperextension were randomly divided into the observation group and the control group with 25 cases in each group. The control group was treated with conventional methods, including Bobath technology, Brunnstrom therapy and motor relearning primarily rehabilitation training. In addition to the conventional methods, patients in the observation group used 8-word bandage to fix knee joint in walking training. Before and after 8 weeks of treatment, Holden walking function classification, 10-meter maximum walking speed and improved Barthel index were adopted to evaluate the walking ability, maximum walking speed and the life ability of the patients. Results There were no significant differences in the scores before treatment between the two groups. The Holden walking function classification, the 10-meter maximum walking speed and the Barthel index scores were significantly improved after 8-week treatment in both two groups (P<0.05), and patient conditions were more significantly improved in the observation group than those of the control group (P<0.05). Conclusion Using 8-word bandage to fix knee joint can significantly improve knee hyperextension in patients with stroke, so as to improve the walking ability and activities of daily living.

Objective To purify pre-ribosome and ribosome of mammalian ceils using continuous sucrose density gradient ultracentrifugation.Methods Continuous sucrose density gradient was established by ultracentrifugation,and the continuous sucrose density gradient of 10％-30％ and 10％-45％ were used to extract the pre-ribosome and ribosome in mammalian cells,respectively.The mammalian cell lysis buffer was added to the established continuous sucrose density gradient.Pre-ribosome and ribosome with different sedimentation coefficients were collected and the A260 absorbance of each sample was measured.Proteins of each sample were extracted to detect the large subunit protein,RPL15 by Western Blot.Results Large subunit ribosomal protein RPL15 exists on 60S of the pre-ribosome,and also on 60S,80S and polyribosome of mature ribosome.Conclusions The continuous sucrose density gradient,which is established by the swing-out rotor,can be used to isolate the pre-ribosome and ribosome of mammalian cells rapidly.This method has the advantages of good separation effect and simple operation,which provides a good method for rapid and large amount preparation and separation of various kinds of ribosomes.

Objective To investigate the process that chromosome kinesin KIF4A promote cisplatin resistance in lung cancer cells.Methods Reverse transcription PCR (RT-PCR) and Western Blot experiments were performed to analyze the expression of KIF4A in lung cancer cells A549 and cisplatin (DDP) resistant cells A549/ DDP.Cell transfection, RNA interference (RNAi) experiments and thiazolyl blue tetrazolium bromide (MTT) assays were carried out to examine cell proliferation of A549 cells with overexpression of exogenous KIF4A and A549/DDP cells with depletion of endogenous KIF4A after cisplatin treatment.Results Expression of KIF4A in A549/DDP cells was higher than that in A549 cells.With overexpression of exogenous KIF4A, A549 cells displayed drug resistance to cisplatin.On the contrary, depletion of endogenous KIF4A in A549/DDP cells resulted in cisplatin sensitivity.Conclusions Chromosome kinesin KIF4A involves in the regulation of cisplatin resistance in lung cancer cells and KIF4A may be a potential and effective new biological target for treatment of lung cancer cisplatin resistance.

To investigate the therapeutic effect and molecular mechanism of the main flavonoid components of Silybum marianum (S. marianum) on nonalcoholic fatty liver disease (NAFLD), we identified nine flavonoids in S. marianum through TCMSP, PubChem database and corresponding literatures. The potential therapeutic targets of NAFLD were predicted by SwissTargetPrediction, GeneCards and Venny 2.1.0 platform, while the protein-protein interaction (PPI) network of potential targets was analyzed using String platform and Cytoscape software. Then GO and KEGG pathway enrichment analysis were performed using David 6.8 database, followed by molecular docking verification using AutoDock software. In vitro, components with higher degree value in the "components-targets-pathway" network were chosen for further analysis. L02 cells were used to establish lipid accumulation model and treated with different components. Furthermore, the effects of four pure active compounds from S. marianum on lipid accumulation in hepatocytes were analyzed by oil red O staining. The results showed that the main nine flavonoids extracted from S. marianum contained 24 potential NAFLD targets. Several critical pathways closely related to NAFLD process were identified by GO and KEGG enrichment analysis, including phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) pathway, type 2 diabetes pathway, tumor necrosis factor (TNF) pathway and insulin resistance pathway. The results of molecular docking further indicated that the core components displayed strong binding abilities with key targets respectively, and silandrin showed better binding activity as compared to other components. The results obtained from L02 cells showed that the lipid accumulation was reduced by treatment with isosilybin A, isosilybin B, silydianin and silychristin, while the activity of isosilybin B was better than that of isosilybin A. Taken together, we concluded that the main flavone components of S. marianum could improve lipid accumulation through multiple signaling pathway in hepatocytes, and this could be a potential new strategy for the treatment of NAFLD.

OBJECTIVETo investigate the effects of emodin on apoptosis induction and proliferation inhibition in human apoptosis and on c-myc protein and mRNA expression in human myeloid leukemia cell line HL-60 cells.

METHODSHL-60 cells were exposed to emodin at different dosages. Growth inhibition was detected by MTT assay and colony formation assay, and cell apoptosis by flow cytometry, TUNEL labeling method, DNA fragmentation and MitoCapture apoptosis detection. The expression of c-myc was detected by RT-PCR and Western-blot.

RESULTSEmodin remarkably inhibited the cell proliferation, with an IC(50) value of 20 micromol/L. HL-60 cells apoptosis could be efficiently induced by emodin in a dose dependent manner. The c-myc protein and mRNA expressions on HL-60 cells were decreased after emodin treatment.

CONCLUSIONEmodin could efficiently induce growth inhibition and apoptosis in HL-60 cells. c-myc may be involved in this process.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Emodin ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; physiology ; RNA, Messenger ; genetics

OBJECTIVETo evaluate the outcome of total knee arthroplasty with or without resurfacing of the patella with particular attention to knee score, knee function score and incidence of postoperative anterior knee pain, providing the basis for the choice of surgical procedure.

METHODSCNKI, PubMed, ScienceDirect, Highwire and other databases were searched for the randomized controlled trials relevant to the patellar with or without replacement in total knee replacement arthroplasty between 1998 and 2010, evaluating of the methodological quality of included studies and extracting valid data.

RESULTSThe 80 citations were identified as related to patellar resurfacing during total knee arthroplasty, 13 articles meet all inclusion criteria for this study. The incidence of postoperative anterior knee pain is greater in knees without replaced patellas (RR = 0.78, 95%CI: 0.61 - 0.99, P = 0.04). No differences are observed between the 2 groups for knee score and knee function score. Knee score (WMD = -0.49, 95%CI: -1.79 - 0.81, P = 0.46), knee function score (WMD = 1.10, 95%CI: -1.77 - 3.98, P = 0.45).

CONCLUSIONSThe patella replacement can significantly reduce the incidence of postoperative anterior knee pain. There is no difference in the knee score and knee function score between two groups.

Arthroplasty, Replacement, Knee ; adverse effects ; methods ; Humans ; Knee Joint ; physiopathology ; Pain, Postoperative ; etiology ; Patella ; surgery ; Randomized Controlled Trials as Topic

ObjectiveTo investigate the predictive value of heart rate turbulence(HRT) in patients with diabetes mellitus (DM) after acute myocardial infarction (AMI).MethodsNinety-two AMI patients combined with DM (DM group) and 120 AMI patients without DM (non-DM group) were selected.Turbulence onset (TO) and turbulence slope (TS) were two indexes of HRT.HRT was considered positive when TO was ≥0 and TS was ≤2.5 ms/R-R.The differences in clinical data between HRT-positive and HRT-negative patients were compared.And the related risk factors after AMI were analyzed.ResultsAge,left ventricular ejection fraction (LVEF) level,renal insufficiency,LVEF＜40％,standard deviation of sinus cardiac cycle (R-R interval)(SDNN),heart rate variability (HRV) positiveand HRT indexes (TO,TS) between HRT-positive and HRT-negative patients in DM group had significant differences (P ＜ 0.05 ).Age,LVEF level,SDNN and HRT indexes(TO,TS) between HRT-positive and HRT-negative patients in non-DM group had significant differences(P ＜ 0.05).Multivariate Cox regression analysis showed that renal insufficiency (OR=4.8,95％ CI:1.8 - 10.7,P=0.008) and HRT positive (OR=3.7,95％ CI:1.5 - 8.6,P=0.070) in DM group had statistical significance.And HRT positive in non-DM group had statisticalsignificance(OR=23.0,95％ CI:5.2 ～ 86.0,P ＜ 0.01 ).ConclusionsHRT,an index of dynamic electrocardiogram,can predict the risk in patients with DM or without DM after AMI.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Carbon Disulfide ; adverse effects ; antagonists & inhibitors ; Cells, Cultured ; Endothelium, Vascular ; drug effects ; Humans ; Umbilical Veins ; cytology ; drug effects ; metabolism

Adolescent ; Anesthetics ; therapeutic use ; Humans ; Male ; Polychondritis, Relapsing ; drug therapy