OBJECTIVETo investigate the effect and mechanism of emodin for regulating aquapoin-2 (AQP2) in NRK cells cultured in vitro.

METHODSExperiments on NRK cells cultured with alpha-DMEM medium in vitro were conducted in two steps. (1) Cells were randomly divided into 4 groups: the control group, and the three emodin treated groups treated with different dosages of emodin (5, 10 and 20 mg/L) respectively. After 24 h treatment, the location of AQP2 was decided by indirect immunofluorescene, and the AQP2 protein and mRNA expression levels were detected by Western blot and semiquantive RT-PCR. (2) Cells were randomly divided into 4 groups, the control group, and the three treated groups treated respectively with 10 mg/L 8-Bromo-cAMP, 20 mg/L emodin, and 20 mg/L emodin +10 mg/L 8-Bromo-cAMP. The activity of protein kinase A (PKA) in NRK cells after 24 h treatment was determined with non-radioactive detecting method.

RESULTSAQP2 was located at the cell membrane of NRK cells. Western blot and semiquantitive RT-PCR found that AQP2 protein and mRNA expressions were significantly decreased in NRK cells of groups treated by 10 mg/L and 20 mg/L emodin (P < 0.05). PKA activity determination showed significantly decreased phosphorylation level of PKA in NRK cells of groups treated with 20 mg/L emodin group (P < 0.05).

CONCLUSIONEmodin can inhibit the genetic transcription and the translation of AQP2 gene in NRK cells, which demonstrates that the change of AQP2 expression regulated by emodin may be correlated with the diuresis effect of rhubarb, and it is likely that the regulation is going through PKA signal pathway.

Animals ; Aquaporin 2 ; genetics ; metabolism ; Cell Line ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Emodin ; pharmacology ; Kidney ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Signal Transduction ; drug effects

BACKGROUNDMammalian target of rapamycin (mTOR) is involved in a caspase independent form of programmed cell death called autophagy. The aim of this research was to investigate the effects of rapamycin and 3-methyladenine (3-MA) on autophagy, proliferation, apoptosis, and cell-cycle parameters of rat bone marrow-derived endothelial progenitor cells (EPCs).

METHODSMononuclear cells isolated from rat bone marrow were treated with rapamycin (0.01, 0.1, 1, or 10 µg/L) or 3-MA (1.25, 2.5, 5, or 10 mmol/L) for 24 hours. Expression of the autophagy marker protein LC3-II was analyzed by Western blotting. Apoptosis and cell-cycle progression were analyzed by flow cytometry. Cell proliferation was measured using the MTT assay.

RESULTSRapamycin treatment of EPCs induced apoptosis and autophagy and inhibited proliferation and cell-cycle progression in a dose-dependent manner. Treatment with 5 mmol/L 3-MA promoted cell proliferation; in contrast, treatment with 10 mmol/L 3-MA promoted apoptosis and induced S-phase arrest.

CONCLUSIONSRapamycin treatment of EPCs induced apoptosis and autophagy. Low concentrations of 3-MA had no significant effect on the proliferation and apoptosis of EPCs; The 5 mmol/L group promoted cell proliferation, but had no effect on the apoptosis; the 10 mmol/L group inhibited the proliferation and promoted apoptosis through the cell cycle.

Adenine ; analogs & derivatives ; pharmacology ; Animals ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Rats ; Sirolimus ; pharmacology

Austria ; Carcinoma, Hepatocellular ; therapy ; Europe ; Hepatitis, Viral, Human ; therapy ; Humans ; Internationality ; Liver Cirrhosis ; therapy ; Liver Neoplasms ; therapy ; Liver Transplantation

OBJECTIVETo explore the effect of compound decoction on notoginsenosides in Panax notoginseng.

METHODNotoginsenoside R1, Rg1, Re, Rb1 and pH were used as the parameters to investigate the changes on the content of notoginsenosides in different compound extractions by heating for two hours and their correlation with pH.

RESULTWhen the pH values of solution of P. notoginseng with Fructus ligustri, P. notoginseng with Eupolyphaga seu steleophaga, P. notoginseng with Pheretima asiatica, and Zhitangjiang Fang (free of Hirudo) were rept higher than 5.7, the reserved rate (RR) of notoginsenside were higher than 90%; When the pH values of decoetion of P. notoginseng with Salvia miltiorrhiza, P. notoginseng with Paeonia lactiflora, P. notoginseng with Platycodon grandiflorum, P. notoginseng with Arctium lappa were kept 4.5-5.5, their RR of notoginsenside were 60% - 85%; When the pH values of the decotction of P. notoginseng with Hirudo nipponica was decreased to 3.4, its RR of of notoginsenside was 38.4%; When the pH values of Zhitangjiang Fang extraction was regulated by 0.1% NaOH solution to pH 6. 3, and the RR of notoginsenside increased to 97%.

CONCLUSIONThe pH of other Chinese herbal medicines extraction with P. notoginseng compound is a critical effect on the stability and yields of notoginsensides.

Animals ; Arctium ; chemistry ; Cockroaches ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ginsenosides ; analysis ; Hirudo medicinalis ; chemistry ; Hot Temperature ; Hydrogen-Ion Concentration ; Ligustrum ; chemistry ; Materia Medica ; chemistry ; isolation & purification ; Oligochaeta ; chemistry ; Paeonia ; chemistry ; Panax ; chemistry ; Platycodon ; chemistry ; Salvia miltiorrhiza ; chemistry

Animals ; Cells, Cultured ; Connective Tissue Growth Factor ; Hepatocytes ; cytology ; metabolism ; Immediate-Early Proteins ; biosynthesis ; genetics ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Male ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Rats ; Transfection

OBJECTIVETo explore the role of glutamine in LPS and D-Gal induced acute hepatic injury.

METHODSA total of 61 Wistar rats were randomly divided into three groups: control group, model group and GLN pretreated group. The animal model was established by LPS and D-Gal intraperitoneal injection. GLN at dose of 1 g/kg was intragastrically administrated for 7 d before intraperitoneal injection. To evaluate the hepatic injury, the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were detected by automatic biochemistry analysator. The liver and bowel tissue was observed by lightmicroscope and transmission electron microscope (TEM). The apoptosis of hepatocyte was detected by TUNEL. HPLC-PED was used in the study of intestinal permeability.

RESULTSNo significant differences were noted between ALT, AST, TBIL level, death rate and intestinal permeability (L/M) between model group and GLN pretreated group; In microscope, the confused structure of hepatic injury and inflammatory infiltration were similar between model group and GLN pretreated group. The injury of bowel was not obviously. Compared with the model group, there was better trend in liver and bowel in GLN pretreated group by transmission electron microscope (TEM). The apoptosis index in GLN pretreated group were lower than those in model group.

CONCLUSIONLPS can induce acute liver injury in D-Gal-sensitized rats.Glutamine has't the trend of protecting liver function and intestinal barrier function,decreasing death rates.

Animals ; Apoptosis ; drug effects ; Female ; Glutamine ; administration & dosage ; Injections, Intraperitoneal ; Intestinal Mucosa ; drug effects ; enzymology ; physiology ; Liver ; drug effects ; injuries ; Random Allocation ; Rats ; Rats, Wistar

Objective To study the mlationship between dietary soy isoflavones and blood lipids among residents of 40-65 years old,in Guangzhou.Methods Dietary soy isoflavones and other nutrients intakes were assessed with quantitative food frequency questionnaire(FFQ).Total cholesterol(TC),triglycerides(TG),HDL cholesterol(HDL-C)and LDL cholesterol(LDL-C)in plasma were measured with colorimetry.Results Ranges of dietary soy isoflavones intake among 134 males and 261 females were fxom 0 mg/day to 61.96 mg/day and 0 mg/day to 82.52 mg/day,with means of 11.95 mg/day,14.90 mg/day,respectively.After adjusted for total energy intake and fat percent energy,difiefences of TC,LDL-C in total population and TC in women were statistically significant between groups(P value was 0.002,0.008,0.004,respectively) and dose-effect relationships(P value was <0.001.0.012.0.001,respectively)were observed between dietary soy isoflavones intake and the upper mentioned three indices.Compared with the low-intake group,tbese three indices lowered 7.06%,10.13%and 7.48%,respectively in high-intake group.Critical significance of LDL-C was observed both in women and men between groups.Further controlled for age,BMI and WHR,no obvious change of the results was observed.Conclusion Moderate intakes of soy isoflavone as part of a regular diet seemed to be associated with favorable blood lipid levels.

OBJECTIVESTo detect the loss of heterozygosity (LOH) of circulating DNA in the plasma of patients with hepatocellular carcinoma (HCC), and to assess its potential as a clinical predictive marker.

METHODSThree high-polymorphic microsatellite markers D8S277, D8S298 and D8S1771 located at chromosome 8p were selected to detect LOH in plasma DNA of 62 HCC patients. The associations between LOH and its clinicopathological features, including HBsAg, liver cirrhosis, serum AFP level, tumor size, tumor cell differentiation, and intrahepatic metastasis were also examined.

RESULTSIn plasma DNA of the 62 HCC patients, LOH was found at one or several loci in 36 (58.1%), and heterozygosity at D8S277, D8S298, and D8S1771 loci was 74.2% (46/62), 75.8% (47/62), and 69.4% (43/62), respectively. LOH frequency at D8S277, D8S298 and D8S1771 was 32.6% (15/46), 44.7% (21/47), and 46.5% (20/43), respectively. LOH in plasma DNA was more frequently detected in the patients with intrahepatic cancer metastasis than those without metastasis (62.5 percent vs. 26.1 percent, P < 0.05); however, no statistically significant correlations were observed between LOH at these loci and other clinicopathological features analyzed in this study.

CONCLUSIONSLOH at D8S298 in plasma DNA may be a potential predictive marker of intrahepatic metastatic recurrence after surgical resection of the HCC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; genetics ; Chromosomes, Human, Pair 8 ; DNA ; blood ; Female ; Humans ; Liver Neoplasms ; genetics ; Loss of Heterozygosity ; Male ; Middle Aged

OBJECTIVETo examine the relationship between intercellular adhesion molecule-1 (ICAM-1) gene polymorphism and ischemic stroke (IS) in Chinese Zhuang populations.

METHODSThe K469E polymorphism in the exon 6 of ICAM-1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism analysis and DNA sequencing in 205 patients with IS of Zhuang nationality and in 210 healthy controls, and the serum level of ICAM-1 was determined by enzyme-linked immunosorbent assay.

RESULTSThe IS group showed significantly higher serum levels of ICAM-1 than did the control group (P < 0.01). There was significant difference in frequencies of allele and genotype in K469E polymorphism between IS and control groups, respectively (P < 0.05). The K allele carriers had 1.424 times the risk of suffering from IS as compared with the E allele carriers (OR = 1.424, 95% CI: 1.071 - 1.894); the serum ICAM-1 level of E allele carriers was significantly higher than that of K allele carriers (501.24 +/- 139.56 ng/ml vs 475.17 +/- 118.35 ng/ml, P < 0.01).

CONCLUSIONThere is an association between ICAM-1 gene K469E polymorphism and IS, and E allele may be a genetic risk factor of IS among Guangxi Zhuangs, in which the ICAM-1 E allele carriers may have up-regulated expression of ICAM-1 and hence are at a higher risk of ischemic stroke.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Brain Ischemia ; ethnology ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

OBJECTIVETo investigate the enhanced predictive activity of preoperative plasma osteopontin (OPN) level in combination with intercellular adhesion molecule-1 (ICAM-1) for recurrence and prognosis of patients after resection of hepatocellular carcinoma (HCC).

METHODSA total of 75 patients received liver resection for HCC from August 2001 to December 2001 in authors' institute were enrolled in this study. The preoperative plasma levels of OPN and ICAM-1 were detected by ELISA, and the association of them combination with the recurrence and prognosis of HCC patients was analyzed.

RESULTSOPN and ICAM-1 could be detected in all of the plasma samples of the tested patients. A significantly higher OPN level and ICAM-1 level were found in plasma of patients who were found to have HCC recurrence during the follow-up time compared with those without recurrence (210.40 vs. 154.86 ng/ml, P = 0.001; 1011.23 vs. 747.49 ng/ml, P = 0.027). A significant difference of OS and DFS were found in different subgroups with higher or lower level of OPN (625 vs. 808 days, P = 0.0006; 433 vs. 674 days, P = 0.0003); and a similar situation was found in patients of high- and low- ICAM-1 levels (651 vs. 794 days, P = 0.0269; 489 vs. 642 days, P = 0.0248). The 2-year recurrence rates of the patients with higher and lower plasma levels of both OPN and ICAM-1 were 87.50% and 28.00% (P < 0.001), respectively; and the 2-year OS rates were 37.50% and 88.00% (P = 0.001), and the 2-year DFS rates were 12.50%, and 76.00 (P = 0.001), respectively.

CONCLUSIONSThe evaluation of preoperative plasma level of OPN or ICAM-1 may be helpful to predict the recurrence and prognosis of HCC patients in advance. The assessment of OPN level in combination with ICAM-1 could stratify patients into groups with different potentials of HCC recurrence and different outcomes more accurately than OPN or ICAM-1 individually.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Liver Neoplasms ; diagnosis ; Male ; Middle Aged ; Osteopontin ; blood ; Prognosis