A PCR assay used for detection and quantification of Neotyphodium lolii in tissues of the grass Lolium perenne L. was developed. Compared with the Microscopic analysis of stained tissues, this method was more accurate. For quantitative applications, real time PCR was used to quantify Neotyphodium lolii in Lolium perenne L.. The results showed that there were some variations between different individuals, but no variance existed between different tillers from the same plant. Thus it can be concluded that fungal content in the plant is not only associated with species/variety specific, but also with the genotype of the host plant as well.

Breast Neoplasms ; chemistry ; genetics ; Chromogenic Compounds ; Female ; Genes, erbB-2 ; Humans ; In Situ Hybridization ; methods ; Receptor, ErbB-2 ; analysis

Photothermal therapy (PTT) has attracted significant attention due to minimal side effects and high treatment specificity. However, it often requires very high temperature to achieve complete tumor ablation under a single PTT. Such high temperature brings obvious thermal damage and inflammatory response to the body, affecting the therapeutic effect. In recent years, nitric oxide (NO) has been used to significantly inhibit tumor growth and enhance the sensitivity of tumor cells of temperature and drugs, thus enhancing the therapeutic effect. However, compounds as NO donors often have some disadvantages such as poor biocompatibility and untargeted delivery, etc., therefore, this medical application based on NO therapy is limited. In conclusion, the organic combination of NO donors and photothermal agents (PTAs) is expected to overcome the shortcomings of single therapy and achieve the antitumor effect of "1 + 1 > 2". In view of the rapid development of NO combining with PTT in tumor therapy, this review firstly introduces the antitumor mechanisms of different types of NO donors. Then the treatment strategy based on NO combined with PTT is discussed. Finally, the prospects and challenges of this combination therapy strategy in the clinical treatment of cancer are discussed.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

Background Although Leber hereditary optic neuropathy (LHON) and optic neuritis have different causes and managements,their clinical manifestations are difficult to be distinguished.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR) is a high flux,simple,rapid and specific detecting technology,so establishing a specific diagnosis method of LHON with RTFQ-PCR has a practical significance.Objective Purpose of the present study was to establish a real-time Taqman probe for mitochondrial DNA (mtDNA)11778G＞A mutation in LHON patients.Methods Primers and Taqman probe for mtDNA 11778G＞A mutation were designed based on mtDNA complete geneme.Eighty-four patients with LHON were selected from the LHON DNA bank of Molecular Biology Laboratory,Henan Eye Institute,and 40 normal physical examinees aged 18-20 years were from Henan People's Hospital.2 ml of periphery blood was collected from each individual.Based on the double-blindness principle,mtDNA 11778G＞A mutation was tested by both Taqman probe and sequencing to check the reliability of real-time Taqman probe.Results The mtDNA 11778G＞A mutation was found in 23 out of 84 patients,and 61 showed a negative result by the technique of real-time Taqman probe.The Ct values of 23 patients with mtDNA 11778G＞A mutation were 22.993 ±0.708,but those of 5 normal controls were 0.These findings showed a consistent rate of 100％ with the sequencing results.In addition,both the false positive rate and the false negative rate were zero.Conclusions Real-time Taqman probe technique is an accurate,convenient,sensitive,specific and intuitionistic method for the diagnosis of mtDNA 11778G＞A mutation in LHON patients.It is feasible and suitable to screen the LHON patients with mtDNA 11778G＞A mutation in a large scale.

Objective To learn the main pathogenic microorganisms,and to acquire the data of pathogens composition anddrug resistance of infant diarrhea disease in Wenzhou City.Methods The diarrheal stool specimens of <5 years oldchildren were collected from outpatients and inpatients in children′s hospital and were cultured and detected.Results Atotal of 201 strains pathogenic bacteria were separated from 51 7 clinical specimens,and the rate of total detection is38.88%,of which included 74 strains intestinal pathogenic bacteria and 1 27 strains diarrhea virus,with the rate of detection1 4.31 % and 24.56%,respectively.Among intestinal pathogenic bacteria,the E.coli was the most common bacteria(36strains,48.65%),followed by salmonella(31 strains,41 .89%).Diarrhea virus included norovirus type 2 (56 strains,44.09%),norovirus type 1 (24 strains,1 8.90%),A group rotavirus (56 strains,31 .50%)and goblet virus(7 strains,5.51 %).The resistance rate of Salmonella to ampicillin was 64.52%,and that to ampicillin,tetracycline,compound newMing were more than 70.00%.The resistance rate of Rifampicin.Novobiocin in all bacteria reached 1 00%.Conclusion E.coli and salmonella epidemic strains were the common pathogenic bacteria in infectious diarrhea among infants inWenzhou City.The predominant viruses were norovirus and rotavirus group A.The drug resistance of various pathogenicbacteria was different,and active surveillance should be strengthened.

5-Aminolevulinate Synthetase/genetics* ; Anemia, Sideroblastic/genetics* ; Genetic Diseases, X-Linked/genetics* ; Humans ; Mutation

OBJECTIVE Non-alcoholic fatty liver disease(NAFLD) encompasses a series of patho-logic changes ranging from steatosis to steatohepatitis,which may progress to cirrhosis and hepatocel-lular carcinoma.The purpose of this study was to determine whether Ganoderma lucidum polysaccha-ride peptide (GLPP) has therapeutic effect on NAFLD. METHODS ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic path-ways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blotting. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD. RESULTS GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, liver dysfunction and liver insulin resistance. Pathways of glycerophospholipid metabolism, fatty acid metabolism and primary bile acid biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1,CYP8B1,FXR,SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited fatty acid synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepato-cytes induced by oleic acid and palmitic acid. CONCLUSION GLPP significantly improves NAFLD via regulating bile acid synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits fatty acid synthesis,indicating that GLPP might be developed as a therapeutic drug for NAFLD.

OBJECTIVETo study the survival and prognostic implication in surgically resected satellite-nodule T4 (T4 satellite) non-small cell lung cancer (NSCLC).

METHODSFrom January 1995 to March 2005, the complete resection was performed to 42 patients with NSCLC who were postoperatively identified as pathologic-stage T4 satellite. Survival and associations between clinicopathological parameters and prognosis were analyzed. Thirty-two patients with pathologic stage local-invasion T4 (T4 invasion) NSCLC who underwent resection at the same time were also analyzed.

RESULTSThe 1-, 3- and 5-year survival was 76.2%, 57.1% and 46.0% for patients with T4 satellite, while 62.3%, 31.5% and 20.0% for patients with T4 invasion. There was a significant higher survival in T4 satellite group when compared to that in T4 invasion group (P < 0.05). Furthermore, patients with T4 satellite N0M0 got a better survival than those with T4 satellite N1-2M0, T4 invasion N0M0 and T4 invasion N1 -2M0 (P < 0.05). For patients with T4 satellite, univariate analysis showed that histology, main tumor size, lymph node status and adjuvant chemotherapy were linked with survival, while main tumor size, lymph node status and adjuvant chemotherapy served as the independent prognostic factors with multivariate analysis.

CONCLUSIONSPatients with completely resected T4 satellite NSCLC have a better prognosis than those with T4 invasion. Main tumor size over 3 cm, lymph node metastasis or no adjuvant chemotherapy means an unfavorable prognosis.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Female ; Humans ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; Prognosis ; Retrospective Studies ; Survival Analysis

OBJECTIVETo investigate the oxidative damage to lung tissue and peripherial blood in PM2.5-treated rats.

METHODSPM2.5 samples were collected using an auto-sampling instrument in summer and winter. Treated samples were endotracheally instilled into rats. Activity of reduced glutathione peroxidase (GSH-Px) and concentration of malondialdehyde (MDA) were used as oxidative damage biomarkers of lung tissue and peripheral blood detected with the biochemical method. DNA migration length (microm) and rate of tail were used as DNA damage biomarkers of lung tissue and peripheral blood detected with the biochemical method.

RESULTSThe activity of GSH-Px and the concentration of MDA in lung tissue significantly decreased after exposure to PM2.5 for 7-14 days. In peripheral blood, the concentration of MDA decreased, but the activity of GSH-Px increased 7 and 14 days after experiments. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood. The DNA migration length (microm) and rate of tail in lung tissue and peripheral blood significantly increased 7 and 14 days after exposure to PM2.5. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood.

CONCLUSIONPM2.5 has a definite oxidative effect on lung tissue and peripheral blood. The activity of GSH-Px and the concentration of MDA are valuable biomarkers of oxidative lung tissue damage induced by PM2.5. The DNA migration length (microm) and rate of tail are simple and valuable biomarkers of PM2.5-induced DNA damage in lung tissues and peripheral blood. The degree of DNA damage in peripheral blood can predict the degree of DNA damage in lung tissue.

Animals ; DNA Damage ; drug effects ; Drug Administration Routes ; Drug Administration Schedule ; Lung ; drug effects ; pathology ; Lung Diseases ; blood ; chemically induced ; pathology ; Male ; Oxidative Stress ; Particle Size ; Particulate Matter ; administration & dosage ; toxicity ; Rats ; Rats, Wistar ; Seasons