Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.

[Objective] To search a suitable preoperative intestinal cleansing method for the operation of colon and rectum. [Methods] A randomized controlled trial was applied in 70 cases before colic and rectal operations. Group A was give:n decoction of Folium Cassiae for oral use and saline enema and Group B was treated with saline enema only. The effect on intestinal cleansing was observed during the operation. [Results] In Group A, 26 cases were remarkably effective in intestinal cleansing, 8 effective, and 1 ineffective and 14, 16 and 5 in Group B respectively ( P

OBJECTIVETo study whether total saponins of Panax notoginseng (PNS) can prevent renal interstitial fibrosis occurrence through blocking the IL-1alpha induced tubular epithelial cell and decrease the secretion of extracellular matrix (ECM).

METHODSNormal rats' tubular epithelial cells NRK52E were cultured in vitro, their morphological changes were observed by inverted phase contrast microscope and scanning electron microscope. Flow cytometry technique and immuno-histochemical method was used to detect the expression of alpha-smooth muscle actin (alpha-SMA), and ELISA was used to quantitatively detect the fibronectin (FN) in the supernatant

RESULTSIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, showing hypertrophy of cells elongated and fusiform-shaped, with significantly enhanced expression of alpha-SMA and increased secretion of FN (P<0.05). After adding PNS of different concentrations, the morphology of cells restored close to normal tubular epithelial cells, with the increase of alpha-SMA expression and FN secretion significantly inhibited (P<0.05) in dose-dependent manner (P<0.05). But addition of different dosages of PNS alone showed no effect on tubular cells.

CONCLUSIONIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, promote the deposition of ECM component FN. PNS could inhibit IL-1alpha induced the transdifferentiation of NRK52E and secretion of ECM, therefore, PNS could be taken as a new drug for prevention and treatment of renal interstitial fibrosis and terminal stage of renal diseases.

Actins ; analysis ; Animals ; Cell Differentiation ; drug effects ; Cell Line ; Epithelial Cells ; cytology ; Fibronectins ; analysis ; Fibrosis ; prevention & control ; Interleukin-1 ; pharmacology ; Kidney ; pathology ; Kidney Tubules ; cytology ; Panax ; chemistry ; Rats ; Saponins ; isolation & purification ; pharmacology

Humans ; Patient Compliance ; Practice Guidelines as Topic ; Pulmonary Disease, Chronic Obstructive ; diagnosis ; etiology ; mortality ; therapy ; Survival Rate

Animals ; Asthma ; drug therapy ; immunology ; pathology ; Cyclohexanecarboxylic Acids ; therapeutic use ; Cytokines ; physiology ; Eosinophils ; physiology ; Genetic Therapy ; Humans ; Inflammation ; pathology ; Matrix Metalloproteinase 2 ; physiology ; Matrix Metalloproteinase 9 ; physiology ; Nitriles ; Quinolines ; therapeutic use ; Signal Transduction

OBJECTIVETo determine butylidenephthalide in Ligusticum Chuanxiong with RP-HPLC.

METHODThe sample was extracted with methanol using sonication. The ESTD was used to quantify butylidenephthalide. HPLC separation was carried out in a Hypersil ODS columm (4.6 mm x 150 mm, 5 microm) , eluted at 1 mL x min(-1) with methanol-5% isopropyl alcohol (60: 40) at 25 degrees C. The detection wavelength was 230 nm.

RESULTThe linear range was 0.07-0.7 microg for butylidenephthalide. The average recovery was 95.3%, and RSD was 2.3% (n =6).

CONCLUSIONThis method was simple and could be used to determine butylidenephthalide with satisfactory accuracy and reproducibility.

China ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Light ; Ligusticum ; chemistry ; Phthalic Anhydrides ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Rhizome ; chemistry ; Scattering, Radiation

Objective To study the alteration of Th17 cells and related molecules in patients with Hashimoto's thyroiditis (HT).Methods Th17 cells were determined by flow cytometry.Real-time PCR method was applied to detect the expression of the orphan nuclear receptor RORγt and interleukin(IL) -17.Serum IL-6 and IL-23 were detected by ELISA method.Results ( 1 ) Compared with healthy controls,the frequency of Th17 cells [ (0.75± 0.79) ％ vs ( 0.28 ± 0.23 )％ ] and expression of RORγt ( 0.30 ± 0.38 vs 0.04 ± 0.02,both P ＜ 0.05 ) were significantly increased in HT patients.( 2 ) The levels of IL-6 and IL23 in HT patients were higher than those in healthy controls ( 3.66 ± 4.70 vs 0.47 ± 1.11,154.7 ± 75.81 vs 80.65 ± 61.41,both P＜0.05 ).( 3 ) A positive correlation between Th17 cells and serum TgAb was revealed in HT patients ( r =0.848 4,P =0.007 7 ).(4) The results of PCR showed that IL-17 and RORγt expressed in thyroid tissues of patients HT.Conclusion An increased frequency of Thl7 cells was found in HT patients,implying that this cell subset may play an important role in the pathogenesis of autoimmune thyroid disease.

AIM: To examine T-lymphocyte rDNA transcription activity in peripheral blood of patients with gastrointestinal maliganant tumor and to clarify its clinical significance.METHODS: T-lymphocyte rDNA transcription activity in peripheral blood of 48 cases of patients with stomach-intestine tract malignant tumor were measured.RESULTS: Before surgery, the T-lymphocyte rDNA transcviption activity was obviously lower than that after surgery, also lower than that of the normal control( P

OBJECTIVETo observe dynamically the Fractalkine (FKN) expression in lung tissue of rats with lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the effect of Shenqi Fuzheng Injection (SFI) on it.

METHODSRat model of ALI was established by intravenous injection of 4 mg/kg of LPS. Forty-two Wistar rats were randomly divided into 7 groups, the normal group, the model group and the SFI group, the latter two were separated respectively into three time phases (the 1 h, 2 h and 4 h after modeling ) groups, 6 rats in each group. Pathological changes and wet/dry weight ratio (W/D) of lung were observed, serum TNF-alpha and FKN mRNA expression in the lung tissue were examined by ELISA and RT-PCR respectively.

RESULTSSevere pathological changes of lung presented in the model groups of all three time phases with a higher W/D ratio, as well as increased serum TNF-alpha level and FKN mRNA expression in lung tissue. The peak of abnormality of serum TNF-alpha level and FKN mRNA expression was shown in the 2 h time phase group. All the above-mentioned abnormal changes were alleviated after treatment in the SFI group (P<0.05). In addition, the level of FKN mRNA expression was found to be positively correlated to the serum TNF-alpha concentration.

CONCLUSIONSFI treatment in early stage could relieve the pathological changes and edema in lung tissue, decrease serum TNF-alpha and down-regulate FKN mRNA expression, playing a protective role in LPS-induced ALI rats.

Animals ; Chemokine CX3CL1 ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Injections ; Lipopolysaccharides ; Lung ; drug effects ; metabolism ; pathology ; Lung Diseases ; blood ; chemically induced ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; blood

The coordination and collaboration of many links are required to achieve the successful development of a high-quality clinical randomized controlled trial, in which, the compliance of the researchers affects the ultimate results and quality of the study since it is responsible for the safety and benefits of subjects in the trials and study quality. Focusing on the researchers' compliance, it is believed that the improvements in the researchers' compliance are very significant in the implementation of study scheme. At present, commonly, the researchers are of the low compliance, manifest as failure to obey the study scheme (such as inclusive criteria, enrollment sequence), failure to obey the rules of operation and data collection, failure to store the original documents but to modify the data, failure to report timely adverse events, etc. In view of clinical research monitors, the reasons on the low compliance are analyzed and the solutions are proposed.