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

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

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

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

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 clarify three-grade criteria of curative resection for primary liver cancer (PLC) and evaluate their clinical significance.

METHODSCriteria of curative resection of PLC were summed up to three grades. Grade I: complete removal of all gross tumors with no residual tumor at the excision margin. Grade II: on the basis of Grade I, there was no extrahepatic metastasis, no hilar lymph node metastasis, no tumor thrombus in the main trunks and their primary tributaries of the portal vein, common hepatic duct, hepatic vein and vena cava inferior, and the tumor was not more than two in number. Grade III: in addition to the above criteria, AFP dropped to normal level (in patients with elevated AFP before surgery) within 2 months after operation, and no residual tumor upon diagnostic imaging. A total of 354 cases with PLC who had their liver resected was reviewed. Patients in each grade were divided into two portions depending on whether the treatment was curative or palliative.

RESULTSThe survival of patients receiving curative treatment was better than those receiving palliative treatment (P < 0.01). This was true for patients whose treatment belonged to anyone of the three-grade criteria. The survival was improved along with the promotion of curative criteria used. The 5-year survival rate of Grade I, II and III patients undergone curative resection was 43.2%, 51.2% and 64.4%, respectively (P < 0.01).

CONCLUSION1. The three-grade criteria may be used for judging the radicality of tumor resection for PLC. 2. The more stringent the criteria used, the better the survival would be. 3. Adopting high-grade criteria to select cases, to guide operation and postoperative follow-up would improve the results of liver resection for PLC.

Female ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; mortality ; surgery ; Male ; Middle Aged ; Survival Rate

OBJECTIVETo evaluate and analyze mechanism of Pizhen for the treatment of pain in patients with knee osteoarthritis.

METHODSEighty-five patients (85 knees) were divided into two groups according to random number table method. Among 35 patients in the experimental group, 6 patients were male and 29 patients were female, 1 patient lost follow-up, the average age was (56.65 +/- 4.89) years, and the patients were treated with Pizhen. Among 50 patients in the control group,there were 6 males and 44 females, the average age was (57.54 +/- 4.91) years, and the patients were treated with intra-articular injection with sodium hyaluronate. The VAS score and the data of algometer from the most obvious tenderness around the knee were used to evaluate the clinical effects.

RESULTSThere were significant differences of scores in the experimental group before and after treatment (P<0.01). There were significant differences of scores in the control group before and after treatment (P<0.01). After treatment, there were no statistical differences of VAS score and the data of algometer between experimental and control groups ( P>0.01). There was negative correlation between VAS score and the data of algometer.

CONCLUSIONThe Pizhen is an effective method to treat pain in patients with knee osteoarthritis by relaxing tendon of soft tissue around pain point of knee joint, and relaxing compress and stimulation of nerves.

Acupuncture Analgesia ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; therapy ; Pain Measurement

Rhubarb is well-known for its cathartic effect, and this cathartic effect, which is closely correlated with "whter" of traditional Chinese medicine (TCM), is brought into play in colon. Recent researches about the relation between formation and effects have identified that the anthraquinone glycosides with 1,8-dio-hydroxy and without hydroxyl in the 2, 3, 6, 7 location, such as emodin, rhein, chrysophanol, et al, can bring about fairly obvious effects of "Watery Diarrhea". Aquaporins (AQPs) are expressed abundantly in colonic epithelial cells, and the abnormal expression of AQPs can lead to the less absorption of water in colon and/or the more secretion of intestinal juice, which suggest that AQPs might be one kind of the effector molecules, which some drugs playing pharmacologic actions in colon depend on. This assumption provides a novel field of vision. Is this "Watery Diarrhea" effect induced by rhubarb concerned with the location alteration or the expression change of AQPs. We deduce that the regulative effects of AQPs by rhubarb in colon might provide a new pharmacologic explation about the cathartic effect through the exploration of TCM and Chinese herbal drugs, with TCM theory and the analysis of data about efficiency and pharmacologic researches of rhubarb and the researches of AQPs. This deduction might be used to reveal why rhubarb can bring about multi-efficiency.
Cathartics ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Rheum ; chemistry