OBJECTIVETo discuss the intervention effect of ligustrazine on ox-LDL-induced foam cells from the perspective of reverse cholesterol transport.

METHODRAW264.7 cultured in vitro was induced with 20 mg x L(-1) ox-LDL to establish the foam cell model, and intervened with ligustrazine. The lipid accumulation in cells was observed by the oil red O dyeing. The changes in total cholesterol and cholesterol ester in the cells were detected with the colorimetric method. The fluorescent quantitative PCR and Western blot were used to detect the mRNA expressions of PPARgamma, LXRalpha and ABCA1.

RESULTLigustrazine could reduce total cholesterol and cholesterol ester in foam cells, inhibit the lipid accumulation, and increase the mRNA and protein expressions of PPARgamma, LXRalpha and ABCA1.

CONCLUSIONLigustrazine can promote the reverse cholesterol transport by increasing the gene expressions of PPARgamma, LXRalpha and ABCA1.

ATP Binding Cassette Transporter 1 ; genetics ; metabolism ; Animals ; Biological Transport ; drug effects ; Cell Line ; Cholesterol ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Foam Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Mice ; PPAR gamma ; genetics ; metabolism

This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of skimmin, scopolin and umbelliferone in Saussurea hieracioides. Samples were analyzed on a Wondasil C18-WR column (4.6 mm x 250 mm, 5 microm) with methanol (A) and water containing 0.1% phosphate (B) as mobile phases for gradient elution at a flow rate of 1.0 mL x min(-1). The detection wavelength and column temperature were set at 325 nm and 35 degrees C, respectively, and the sample size was 10 microL. The results showed that skimmin, scopolin and umbelliferone were simultaneously achieved within 40 min under the above conditions. A good linearity was observed in the range of 0.18-5.6 microg (r = 1.000 0), 0.060-1.8 microg (r = 0.999 9), 0.032-0.97 microg (r = 0.999 8) for skimmin, scopolin and umbelliferone, respectively, with the average recoveries of 99.16% (RSD = 0.41%), 100.3% (RSD = 0.79%), 102.2% (RSD = 0.87%). The method is simple, accurate and reproducible and can be used for the quality control of S. hieracioides.
Chromatography, High Pressure Liquid ; Coumarins ; analysis ; Glucosides ; analysis ; Medicine, Tibetan Traditional ; Reproducibility of Results ; Saussurea ; chemistry ; Umbelliferones ; analysis

Objective To investigate the real experiences of nurses participating in the treatment of patients with new respiratory infectious diseases. Method The semi-structural interview method was used to carry out in-depth interviews with 8 nurses who were involved in the whole process of treatments and cares of patients with SARS and H1N1 influenza to understand their real experiences. Result The nurses, who participated in the treatment and care to patients with new respiratory infectious diseases, experienced such emotions of nervousness, excitement, anxiety, fear, exhaustion, obtained support from their families, hospitals, society and meanwhile recognized their lack of professional knowledge and clinical skills. Conclusion The nurses participating in the treatment of patients with new respiratory infectious diseases should be given strengthened professional training , psychological and social support.

Objective To assess the value of three-dimensional contrast-enhanced ultrasonography (3D-CEUS) in evaluation of hepatic arteries variants.Methods Both two-dimensional contrast-enhanced ultrasonography (2D-CEUS) and 3D-CEUS were used to assess 30 patients including living donor candidates and patients with upper abdomen tumors.The reference standard was operation or CTA or DSA or MRA,and the accuracy for detecting hepatic artery variants provided by the two methods was evaluated.Arterial anatomic types were defined by using Michels classification.Results The total accuracy for detecting hepatic artery anatomy types by 2D-CEUS was 40.0% (12/30),while 83.3% (25/30) by 2D-CEUS.For convention anatomy types the accuracy on 2D-CEUS and 3D-CEUS were 40.9%(9/22)and 90.9%(20/22),respectively.The difference was statistically significant (P<0.05).For anatomy variants types the accuracy on 2D-CEUS and 3D-CEUS were 37.5%(3/8)and 62.5%(5/8),respectively.No significant difference between these two methods was observed.Conclusion 3D-CEUS was a new method in diagnosis of hepatic arteries anatomy types with practical clinical value in evaluation of the living liver donors.

Objective To investigate the effect of berberine on insulin resistance induced by free fatty acid in 3T3-L1 adipocytes and the possible molecular mechanism.Methods 3T3-L1 adipocytes were treated with 0.5 mmol/L palmitic acid to induce insulin resistance.Berberine was used for treatment and aspirin for positive control.Glucose oxidase method was employed for measuring the glucose consumption in the medium and 2-deoxy- [~3H]-D-glucose method was used for the determination of glucose uptake.Western blot was used for the determination of IKB kinase(IKK)?SerlS1 phosphorylation,insulin receptor substrance-1(IRS-1)Ser307 phosphorylation,the protein expression of IKK?,IRS-1,phosphatidylinositol 3-kinase(PI-3K)p85 and glucose transporter 4(Glut4).Results After the treatment with 0. 5 mmol/L of palmitic acid for 24 h,glucose consumption by 3T3-L1 adipocytes was decreased by 41%,insulin-stimulated glucose transport was inhibited by 67%,IRS-1 and PI-3K p85 proteins were reduced, and phosphorylations of IKK?Ser181 and IRS-1 Ser307 were induced.The above results were reversed by adding berberine or aspirin.But Glut4 and IKK?protein abundance was not changed during this study.Conclusion Berberine significantly improves insulin resistance induced by free fatty acid in 3T3-L1 adipocytes via inhibiting IKK?serine phosphorylation.

OBJECTIVETo establish a method for studying efficacious materials of traditional Chinese medicines from an overall perspective.

METHODCarthamus tinctorius was taken the example. Its major components were depleted by preparing liquid chromatography. Afterwards, the samples with major components depleted were evaluated for their antioxidant effect, so as to compare and analyze the major efficacious materials of C. tinctorius with antioxidant activity and the contributions.

RESULTSeven major components were depleted from C. tinctorius samples, and six of them were identified with MS data and control comparison. After all of the samples including depleted materials are compared and evaluated for their antioxidant effect, the findings showed that hydroxysafflor yellow A, anhydrosafflor yellow B and 6-hydroxykaempferol-3, 6-di-O-glucoside-7-O-glucuronide were the major efficacious materials.

CONCLUSIONThis study explored a novel and effective method for studying efficacious materials of traditional Chinese medicines. Through this method, we could explain the direct and indirect contributions of different components to the efficacy of traditional Chinese medicines, and make the efficacious material expression of traditional Chinese medicines clearer.

Alkalies ; chemistry ; Antioxidants ; chemistry ; Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Mass Spectrometry

Osteoarthritis (OA) is a complex chronic progressive disease attacked by biological and mechanical factors and a result from the anabolic and catabolic imbalance in chondrocyte, subchondral bone and extracellular matrix(ECM). Etiology and pathological of OA are not yet entirely clear. The degradation and destruction of collagen II caused by matrix metalloproteinase -13 (MMP-13) is considered the core factor in the occurrence and development of OA. The research of MMP-13 inhibitor provide ideas and methods for the treatment of OA. In this article,the role and determination of MMP-13 in OA and the development prospect of MMP-13 inhibitor in the treatment of OA research progress were reviewed.
Animals ; Collagen ; metabolism ; Humans ; Matrix Metalloproteinase 13 ; analysis ; physiology ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Osteoarthritis ; drug therapy ; etiology

Oral submucous fibrosis (OSF) is a potentially malignant disorder that is characterized by a progressive fibrosis in the oral submucosa. Arecoline, an alkaloid compound of the areca nut, is reported to be a major aetiological factor in the development of OSF. Low-power laser irradiation (LPLI) has been reported to be beneficial in fibrosis prevention in different damaged organs. The aim of this study was to investigate the potential therapeutic effects of LPLI on arecoline-induced fibrosis. Arecoline-stimulated human gingival fibroblasts (HGFs) were treated with or without LPLI. The expression levels of the fibrotic marker genes alpha-smooth muscle actin (α-SMA) and connective tissue growth factor (CTGF/CCN2) were analysed by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) and western blots. In addition, the transcriptional activity of CCN2 was further determined by a reporter assay. The results indicated that arecoline increased the messenger RNA and protein expression of CCN2 and α-SMA in HGF. Interestingly, both LPLI and forskolin, an adenylyl cyclase activator, reduced the expression of arecoline-mediated fibrotic marker genes and inhibited the transcriptional activity of CCN2. Moreover, pretreatment with SQ22536, an adenylyl cyclase inhibitor, blocked LPLI's inhibition of the expression of arecoline-mediated fibrotic marker genes. Our data suggest that LPLI may inhibit the expression of arecoline-mediated fibrotic marker genes via the cAMP signalling pathway.

OBJECTIVETo explore the prediction of the site for microsurgical vasoepididymostomy (VE) in the treatment of epididymal obstructive azoospermia (OA).

METHODSThis study involved 56 infertile men with confirmed OA whose obstruction was suspected to be in the epididymis. Based on their medical history and results of preoperative physical examination and ultrasonography, we predicted the sites for VE. We performed surgical scrotal exploration for the status of epididymal obstruction, conducted palpation and microscopic observation for the epididymal tubules to be anastomosed, and finally decided on the sites for VE by making sure of the presence of motile sperm in the epididymal fluid of the patients. After surgery, we followed up the patients for the rate of pregnancy.

RESULTSAll the patients received bilateral scrotal ultrasonography and surgical scrotal exploration, totaling 112 procedures, including 98 VE procedures. The accuracy rate of the predicted sites for VE was 80.5% (153/190) by medical history and physical examination, 80.3% (90/112) based on the results of ultrasonography, and 87.4% (90/103) according to the first selected epididymal tubules. Of the 28 patients followed up for more than 12 months, motile sperm were found in 19 (67.9% ) at 2 to 12 months and spontaneous pregnancies were achieved in 10 (35.7%), all with the anastomotic sites in the corpus or cauda.

CONCLUSIONMedical history and physical examination contribute to the selection of anastomotic sites and non-invasive scrotal ultrasonography is effective and practical for positioning epididymal obstruction. The epididymal tubules with motile sperm for anastomosis could be easily obtained from the most dilated ones in indurated epididymides.

Azoospermia ; surgery ; Body Fluids ; Epididymis ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Microsurgery ; methods ; Pregnancy ; Pregnancy Rate ; Scrotum ; diagnostic imaging ; Ultrasonography ; Vas Deferens ; diagnostic imaging ; surgery

OBJECTIVETo develop HDV as a vehicle to deliver hammerhead ribozyme into hepatocytes, the effects of modified HDV was assessed on the activity of embedded hammerhead ribozyme in vitro and in vivo.

METHODSIn vitro activity of ribozyme or HDV-driven ribozyme was assessed by incubating with the [alpha-32 P]-ATP labeled HBV RNA substrates at different temperature. Huh-7 cells were cotransfected with ribozyme or HDV-ribozyme chimera and HBV genome, by which inhibition of ribozymes on HBV transcription in vivo were examined.

RESULTSThe results indicated that both temperature and secondary structure influenced the cleavage activity of HDV-driven ribozyme significantly. When the factors were eliminated, the HDV-driven ribozyme could act as well as its counterpart naked ribozyme. While in cultured cells the HDV-driven ribozyme had higher inhibition to HBV gene expression than that of ribozyme alone.

CONCLUSIONThe results demonstrated that HDV may weaken the activity of embedded ribozyme in vitro, but make it enhanced in cultured cells. Thus, this study could provide a useful evidence to develop HDV as vector for liver-special delivery of ribozyme to against chronic HBV infection.

Base Sequence ; Cell Line, Tumor ; Genome, Viral ; Hepatitis Delta Virus ; enzymology ; genetics ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Protein Structure, Secondary ; RNA, Catalytic ; chemistry ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Structure-Activity Relationship ; Substrate Specificity ; Temperature ; Transfection