Objective: Based on optimized method of DNA ligase chain reaction in medium/low throughput genotyping, we assessed the relationship between NOD-like receptor genes NOD1, NOD2 and coronary artery disease (CAD) occurrence. Methods: A multiplex PCR was conducted to enrich DNA template; probe design, annealing temperature, time and number of circulation of PCR were opfimizecl for allele specific ligation; allele specific products were identified by fluorescence PCR and capillary electrophoresis; the accuracy was verified by Sanger sequencing. Single nucleotide polymorphisms (SNPs) on NOD1 gene and NOD2 gene were examined in 1555 CAD patients and 1887 control subjects; the relationship between SNPs and CAD occurrence was studied. Results: Based on optimized PCR condition and allele specific probe design, 30 allele loci genotyping can be genotyped by 10ng DNA template at one time. Association study presented that rs751271 and rs1861759 on NOD2 gene were related to non-hypertensive CAD, allP<0.05; with Bonferroni correction, such correlation was still significant, allP<0.05. Conclusion: We optimized DNA ligase chain reaction and established a novel high accuracy, low cost method for the demand of medium/low throughput genotyping in clinical molecular diagnosis. With this method, we identified that rs1861759 and rs751271 on NOD2 gene were associated with non-hypertensive CAD.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Hippocampus ; cytology ; Neurons ; cytology ; drug effects ; Propofol ; pharmacology ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats

BACKGROUND:Tissue inhibitor of matrix metaloproteinase 1 (TIMP-1) is the corresponding antagonist of matrix metaloproteinase 13 (MMP-13), and their balance between expression and functional activity exerts an important role in the metabolic state of the extracelular matrix. During the development of osteoarthritis, however, TIMP-1 and MMP-13 expressions and their expression ratio show unclear changes in DH guinea pigs.
OBJECTIVE:To explore the expression levels of MMP-13 and TIMP-1 in DH guinea pigs with different ages, and to analyze the relationship between the ratio of MMP-13 to TIMP-1 and the age-dependent degenerative changes in the articular cartilage.
METHODS:Twenty-four female Dunkin Hartley guinea pigs were sacrificed at age of 2, 4, 8, 12 months separately, with six animals at each time point. The knee joints were colected and gross visual appearance of the articular cartilage was observed, then were decalcified and prepared for paraffin sections. VG staining and Mankin score were used to analyze the histological changes. Immunohistochemistry was conducted to assess the expression levels of MMP-13 and TIMP-1 in the cartilage. Integrated absorbance values were used as the quantitive analysis calculated by Image pro-Plus 6.0. Linear regression analysis was done to analysis the relationship between Mankin score and the ratio of MMP-13/TIMP-1. RESULTS AND CONCLUSION:Normal appearance in the articular cartilage was observed in 2-month-old DH guinea pigs, while degenerative changes in the articular cartilage were shown in 4-month-old animals, which became severer with age. Significant difference was found in Mankin score between any two groups (P < 0.05). The ratio of MMP-13 to TIMP-1 increased with age, and the ratio was positively correlated to the Mankin score (P < 0.05). Age-related articular cartilage degeneration occurred in Dunkin Hartley guinea pigs at 4 months of age, and devoloped with age, which is related with the imbanlance of the expression ratio of MMP-13 to TIMP-1.

OBJECTIVE:To establish a method for determining the plasma concentration of paeoniflorin and phillyrin and phar-macokinetic study before and after intragastric administration of Qianliean granules. METHODS:LC-MS/MS method was adopted. The column was Waters C18 with mobile phase consisted of acetonitrile(A)-2 mmol/L ammonium acetate(containing 0.05% formic acid)(B)(0-9 min:15%A→50%A;9-11 min:50%A→90%A;11-17 min:90%A;17-19 min:90%A→15%A;19-20 min:15%A),at the flow rate of 0.6 ml/min;column temperature was 35 ℃ and the volume was 20 μl;quantitative ions were paeoniflorin m/z 525.2 → m/z 449.0,phillyrin m/z 552.3 → m/z 355.3. 7 SD male rats were docked to collect blood 0.5 ml from angular vein 0.25,0.5,0.75,1,1.5,2,3,4,6,8,10,12,24 h after administration Qianliean granule solution 1 g(medicinal materials)/kg to determine the blood concentration of drugs. DAS 2.1.1 software was employed to calculate pharmacokinetic parameters. RE-SULTS:The linear range of paeoniflorin and phillyri were 5.0-2500.0 μg/L(r=0.9979)and 2.0-2000.0 μg/L(r=0.9982),re-spectively;RSD of precision test was less than 5.5%(n=5);the method recovery were 96.0%-104.0% and 92.0%-107.0%,the extration recovery were 71.4%-83.5% and 81.5%-92.3% and RSD of stability test was less than 5.0%(n=3). The pharmacokinet-ic parameters of paeoniflorin and phillyrin were as follows as t1/2 of (2.206 ± 0.631) and (1.355 ± 0.317) h;cmax of (1504.069 ± 620.885) and (79.043 ± 15.568)μg/L;tmax of (1.000 ± 0.250) and (1.214 ± 0.267) h;AUC0-24 h of (4897.645 ± 2207.577) and (263.475±54.795)μg·h/L;CL of(5.025±2.773)and(76.253±13.986)L/(h·kg). CONCLUSIONS:The method is highly sensi-tive,exclusive,simple,accurate and reliable,and can be applied to study the pharmacokinetic characteristics of paeoniflorin and phillyrin in rats in vivo.

Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

Aim To investigate the antioxidant capacity of crocetin and crocin in an in vivo system.Methods Column chromatography was applied to the seperation of crocetin and crocin-1 from gardenia.Crocetin(6.25,12.5 and 25.0 mg·kg~(-1)·d~(-1)) and crocin (18.7,37.5 and 75.0 mg·kg~(-1)·d~(-1)) were orally administered to kunming mice.Then,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total antioxidant capacity(TAOC)and malondialdehyde(MDA)in mice were determined for the comparison of antioxidant activity of crocetin and crocin-1.Results Oral administration of crocetin and crocin for six weeks could enhance SOD of liver and kidney,GSH-Px of liver and TAOC of heart and kidney.In addition,it could decrease MDA of serum in mice.Conclusions The comparison of results suggests the evidence supporting the comparable antioxidant activity of crocetin and crocin.The results of the research also indicate that liver and kidney are two organs targeted for protection concerning endogenous antioxidant among various tissues.

Objective To identify histopathologic correlates for the various MRI appearances of breast fibroadenomas.Methods Thirty-eight fibroadenomas in 33 patients(aged 24—57 years)examined with gadolonium-enhanced MR imaging were observed for signal intensity on T_2-weighted images,contrast enhancement,shape,and internal septation,and these findings were correlated with histopathologic findings.All cases underwent surgery and were proved by pathology.Results(1)The lesion shape was lobular,or round in 34 of 38 fibroadenomas(89.5%).(2)The signal intensity on T_1-weighted images was less than or equal to that of fibroglandular tissue in all cases.The signal intensity on T_2-weighted images was highly varible:high T_2 signal intensity was associated with more myxomatous stromal(mean myxoid-sclerotic index value of 1.9),higher stromal cellularity(mean stromal cellularity index value of 2.2); Fibroadenomas with low T_2 signal intensity had stromal that was nearly uniformly sclerotic(mean myxoid- sclerotic index values of 2.8)and low stromal cellularity(mean stromal-cellularity index value of 1.2). Significant differences were found between these two groups,x~2=11.267 and x~2=10.415(P0.05).The degree of contrast enhancement was proved to be related to ages of patients.The enhancement was more intensely in younger patients.(5)Internal septations were identified within nine of 33 enhancing fibroadenomas (27.3%)and appeared to correlated with collagenous bands at histopatholigic analysis.Conclusions Fibroadenomas demonstrate marked histopathologic variability.The resultant variability in the MR appearance correlated with the degree of myxomatous or sclerotic and stromal cellularity.Lobulation and internal septation,which appear to reflect intrinsic growth patterns of fibroadenomas,may provide more reliable information for distinction.Familiarity with the diagnostic features would facilitate to make the differential diagnosis correctly.

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

Traditional Chinese medicine Polygoni Multiflori Radix is dried roots of Polygonaceae Polygortum multiflorum Thunb. Its clinical application records were first discovered in literatures of the Tang dynasty. The origins, efficacy, toxicity, processing and taboos of Polygoni Multiflori Radix have been discussed in many ancient herb literatures. In recent years, with the increase in the public awareness in health, Polygoni Multiflori Radix admits preparations have been more widely applied in the treatment and prevention of diseases. However, there have been more and more reports about Polygoni Multiflori Radix induced liver injury, the safety of Polygoni Multiflori Radix has increasingly attracted attention of the society. In this paper, the authors summarized and analyzed the toxicity and medication risk factors of Polygoni Multiflori Radix recorded in ancient herb literatures, and proposed that more attention shall be given to the effect of the planting and processing methods on the components and toxicity of Polygoni Multiflori Radix in safety studies, which provides clues for the further studies.
China ; Drugs, Chinese Herbal ; adverse effects ; history ; History, Ancient ; Humans ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; Polygonum ; adverse effects ; Risk Assessment

The renal toxicity and mutagenicity of aristolochic acid (AA) as well as its carcinogenicity on upper urinary tract transitional epithelial cells have been widely known. Since 2003, drug regulatory departments have successively cancelled the quality standards for AA-containing medicines such as Aristolochiae Radix, Aristolochiae Manshuriensis Caulis and Aristolchiae Fangchi Radix, and adopted measures for strengthening regulation and revising package insert or quality standards for other AA-containing medicines, including Aristolochia Cinnabarina Radix, Aristolochiae Fructus, Aristolochiae Mollissimae Herba, in order to control its safety risk. In recent years, domestic and foreign studies on AA have mainly involved action mechanism and clinical performance of AA toxicity, early-stage diagnosis and treatment method. In this paper, authors gave a brief summary and evaluation on risk factors for using AA-containing medicines, and offered measures and suggestions for preventing and controlling AA toxicity.
Animals ; Aristolochia ; adverse effects ; chemistry ; Aristolochic Acids ; analysis ; therapeutic use ; toxicity ; Drug Therapy ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; analysis ; therapeutic use ; toxicity ; Humans