Objective: To investigate the effect of all trans retinoid acid (ATRA) on bone cell proliferation and differentiation. Methods: Newborn rat calvarial osteoblastic cells were isolated and the metabolism of the osteoblastic cells were determined by MTT, Goldens method and immunocytohistologic method. Results: 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 mol/L ATRA could increase osteoblastic cells proliferation after 72 h culture; 10 -5 , 10 -6 , 10 -7 mol/L ATRA could increase ALP activity. The expression of cyclin D 1 was decreased. Conclusion: ATRA stimulates the proliferation and differentiation of rat calvarial osteoblastic cells in vitro. Cell cycle relative proteins may play an important role in control of cell proliferation and differentiation induced by retinoic acid and derivatives.

OBJECTIVETo evaluate the outcomes of T3a prostate cancer with unfavorable prognostic factors treated with permanent interstitial brachytherapy combined with external radiotherapy and hormone therapy.

METHODSFrom January 2003 to December 2008, 38 patients classified as T3a prostate cancer with unfavorable prognostic factors were treated with trimodality therapy (brachytherapy + external radiotherapy + hormone therapy). The prescription dose of brachytherapy and external radiotherapy were 110 Gy and 45 Gy, respectively. The duration of hormone therapy was 2-3 years. The endpoints of this study included biochemical failure-free survival (BFFS), distant metastasis-free survival (DMFS), cancer-specific survival (CSS), and overall survival (OS). Survival curves were calculated using the Kaplan-Meier method. The Log-rank test was used to identify the prognostic predictors for univariate analysis.

RESULTSThe median follow-up was 71 months. The serum pre-treatment prostate-specific antigen (PSA) level ranged from 10.0 to 99.8 ng/ml (mean 56.3 ng/ml), the Gleason score ranged from 5 to 9 (median 8), and the percentage of positive biopsy cores ranged from 10% to 100% (mean 65%). The 5-year BFFS, DMFS, CSS, and OS rates were 44%, 69%, 82%, and 76%, respectively. All biochemical failures occurred within 40 months. The percentage of positive biopsy cores was significantly correlated with BFFS, DMFS, and OS (all P=0.000), and the Gleason score with DMFS (P=0.000) and OS (P=0.001).

CONCLUSIONST3a prostate cancer with unfavorable prognostic factors presents not so optimistic outcome. Hormone therapy should be applied to prolong the biochemical progression-free or metastasis-free survival. The percentage of positive biopsy cores and the Gleason score are significant prognostic factors.

Androgen Antagonists ; therapeutic use ; Brachytherapy ; Combined Modality Therapy ; Gonadotropin-Releasing Hormone ; agonists ; Humans ; Male ; Neoplasm Grading ; Prognosis ; Prostatic Neoplasms ; mortality ; pathology ; therapy ; Treatment Outcome

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

Objective To investigate the correlation between IL‐18 and deep venous thrombosis disease and its clinical significa‐tion .Methods To detect the expression of IL‐18 by ELISA ,we collected the blood samples of DVT patients as the experimental group(n=40) compared to the control group(n=40) and normal group(n=20) .IL‐18 over expression/interference vectors were constructed and transfected human vein endothelial cells ,analyzed by microarray and KEGG Pathway as biology information tech‐nology .Then discuss the association between IL‐18 and DVT .Results Results of ELISA showed that compared with control group and normal group ,the expression of IL‐18 gene in DVT patient were up‐regulated(F=11 .248 ,P<0 .01) .Compared with normal group ,the IL‐18 expression in control group have not been significantly up‐regulated(P>0 .05) .Immunofluorescence detected IL‐18 gene expression in cytoplasm of human umbilical vein endothelial cells (HUVECs) .According to the microarray analysis we found in the IL18‐pCDH‐GFP transfected cells 17 signaling pathways were down‐expressed while 16 signaling pathways were up‐expressed .Compared with normal group cells ,in the IL18‐LMP‐shRNAmir1 transfected cells 23 signaling pathways were down‐ex‐pressed and 9 signaling pathways were up‐expressed .Conclusion Based on the above experimental data ,it is very clear that IL‐18 influenced HUVECs and plays an important role in DVT ,it is possible to predict the diagnosis of DVT and act as candidate molecu‐lar markers .

Antimicrobial peptides, a cluster of small peptides secreted by the majority of creatures, have been demonstrated with activity against a wide range of microorganisms including bacteria, protozoa, yeast, fungi, viruses and even tumor cells. These peptides have some features such as broad spectrum , high effi-cacy and stability, little drug resistance. A lack of new antibiotics combined with emerging multi-drug resis-tance issues demands that new antimicrobial strategies be explored for treating these infections. It has been proposed that the antimicrobial peptides might form the foundation for a new class of clinically useful an-timicrobials. We review the advantages of these molecules in construction features and bioactivity, with the focus on the mechanism and clinical applications.

OBJECTIVETo explore the feasibility of evaluating complete ischemia-reperfusion injury (IRI) of the testis by contrast-enhanced ultrasonography with microbubbles (MB) targeted to P-selectin (MBp) in rabbits.

METHODSWe randomly divided 30 healthy adult rabbits into five groups of equal number (control, 0.5 h IRI, 1 h IRI, 2 h IRI, and 4 h IRI), prepared phospholipid MB and MBp, and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection, we recorded the power of the first frame (F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue.

RESULTSCEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups (P < 0.05), which was (8.34 +/- 1.20) versus (1.87 +/- 0.25) 10(-5) AU at 2 hours, but there was no significant difference between MB and MBp in the control rabbits (0 AU, P > 0.05). Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes, but not in those of the control.

CONCLUSIONContrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.

Animals ; Antibodies ; Disease Models, Animal ; Male ; Microbubbles ; P-Selectin ; immunology ; Rabbits ; Reperfusion Injury ; diagnostic imaging ; Testis ; blood supply ; Ultrasonography

Background and Objective:Thrombospondin-1(TSP1)is an inhibitor of angiogenesis and its promoter hypermethylation has been found resulting in gene silencing in some primary human carcinomas.This study was to investigate the promoter methylation of TSP1 and its correlation with TGF-β1 level and T cell lmmunity in gastric cardia adenocarcinoma(GCA).Methods:Methylation specific polymerase chain reaction(MSP)approach and immunohistochemistry method were used to examine the methylation status of the 5'CpG island and expression of TSP1 protein, respectively.The Ievel of TGF-β1 was measured by ELISA and T cell immunity of GCA by flow cytometry analysis.Results:TSP1 methylation frequency was significantly higher in tumor specimens than in corresponding normal tissues(35.4% vs.3.1%,P<0.001)and significanty higher in Stages Ⅲ and Ⅳ tumor tissues than in Stages Ⅰ and Ⅱ tumor tissues(P<0.05).TSP1 protein expression was significantly lower in the tumor tissues than in corresponding normal tissues (P<0.05)and statistically correlated with its methylation status(P0.05).The function of T cell immunity was significantly different between the GCA patients with hypermethylation of TSP1 and those without methylation of TSP1(P<0.05).Conclusion:Promoter hypermethylation of TSP1 may play an important role in the development of GCA and reflect the biological behaviours of GCA.

OBJECTIVETo probe into the mechanisms of moxibustion and Chinese herbs in delaying aging.

METHODSSixty SD rats were randomly divided into a young control group, an aging group, a moxibustion group, a Chinese herb group and a moxibustion plus Chinese herb group. In the latter 4 groups, aging rat model was established by hypodermic injection of D-galactose. In the course of modeling, the 3 treatment group were treated by mild-warm moxibustion at "Zusanli (ST 36)", "Shenshu (BL 23)" and "Guanyuan (CV 4)" with reinforcing method, stomach perfusion of decoction of Liuwei Dihuang plus Danggui (Angelica) and Danshen (Salvia Miltiorrhiza Bge), and the moxibustion plus the Chinese herbs, respectively. After treatment for 40 days, liver mitochondrial DNA, serum IL-2 and IL-6 contents were detected.

RESULTS(1) The mitochondrial DNA content of liver cells and serum IL-6 level significantly increased (P < 0.05, P < 0.01) and serum IL-2 level significantly decreased (P < 0.05) in the aging model group as compared with those in the young control group. (2) The mitochondrial DNA content of liver cells and serum IL-6 level significantly decreased (P < 0.05, P < 0.01) and serum IL-2 level significantly increased (P < 0.05) in all the treatment group as compared with those in the aging model group.

CONCLUSIONMoxibustion and Chinese herbs function delaying aging though decreasing mitochondrial DNA content of liver cells and serum IL-6 level and increasing serum IL-2 level.

Aging ; drug effects ; immunology ; Animals ; DNA, Mitochondrial ; analysis ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Medicine, Chinese Traditional ; Moxibustion ; Rats ; Rats, Sprague-Dawley

BACKGROUNDCongestive heart failure (CHF) is associated with Cheyne-Stokes respiration (CSR), which may hasten CHF. Adaptive servoventilation (ASV) is a novel method of ventilatory support designed for removal of CSF in CHF patients. This study compares the efficacy of ASV in patients with CHF and CSR with the efficacy of oxygen therapy.

METHODSFourteen patients with CHF and CSR were recruited. During sleep, nasal oxygen therapy and ASV treatment were each performed for two weeks. Comparison before and after each treatment was made for the following items: a) parameters of sleep respiration, sleep structure and quality; b) left ventricle ejection fraction (LVEF) and 6-minute walk distance.

RESULTSCompared with the baseline levels of apnoea hypopnoea index of 34.5 +/- 6.1 before treatment, the apnoea hypopnoea index significantly decreased following oxygen therapy to 27.8 +/- 8.2, P < 0.05 and further reduced following ASV treatment to 6.5 +/- 0.8, P < 0.01. The minimal pulse oxygen saturation markedly increased following oxygen therapy from a baseline of (84.3 +/- 2.6)% to (88.6 +/- 3.7)%, P < 0.05 and further increased following ASV treatment (92.1 +/- 4.9)%, P < 0.01. Stages I + II sleep as percentage of total sleep time decreased from (81.9 +/- 7.1)% to (78.4 +/- 6.7)% following oxygen therapy and further to (72.4 +/- 5.0)% following ASV treatment. Stages III + IV sleep as percentage of total sleep time decreased from (8.4 +/- 5.5)% to (6.0 +/- 3.0)% following oxygen therapy and but increased to (11.9 +/- 5.4)% following ASV treatment. The arousal index of 30.4 +/- 8.1 before treatment significantly decreased following oxygen therapy to 25.6 +/- 5.7, P < 0.05 and further declined following ASV treatment to 18.2 +/- 6.1, P < 0.01. No significant difference was shown in above percentages between day 14 of oxygen therapy and before treatment (P > 0.05). LVEF was significantly higher on day 14 of ASV treatment (37.2 +/- 4.1)% than on day 14 of oxygen therapy (33.2 +/- 5.1)% and before treatment (30.2 +/- 4.6)% (all P < 0.05). Six-minute walk distance was the shortest before treatment (226 +/- 28) m, longer on day 14 of oxygen therapy (289 +/- 26) m, and the longest on day 14 of ASV treatment (341 +/- 27) m (all P < 0.01).

CONCLUSIONASV treatment is of better efficacy and greater clinical significance in improvement of CHF by eliminating CSR than oxygen therapy.

Adult ; Cheyne-Stokes Respiration ; physiopathology ; therapy ; Female ; Heart Failure ; complications ; physiopathology ; Humans ; Male ; Middle Aged ; Oxygen Inhalation Therapy ; Positive-Pressure Respiration ; methods ; Sleep ; physiology ; Stroke Volume ; Ventricular Function, Left

AIMTo study the effect of taurine (Tau) on rabbit cardiomyocyte apoptosis during ischemia/reperfusion (I/R) injury.

METHODSRabbit heart I/R injury was induced by occluding the left anterior descending coronary artery for 45 min and reperfusion for 180 min. taurine (200 mg/kg) was intravenously injected 5 min before heart ischemia. Cardiomyocyte apoptosis was measured by using terminal deoxynucleotidyl transferase--mediated dUTP nick end labeling method (TUNEL), flow cytometry (FCM) and DNA agarose gel electrophoresis.

RESULTSDNA ladder pattern of DNA in myocardium was revealed by agarose gel electrophoresis in I/R group while was not found in Tau + I/R group. Apoptotic cardiomyocytes were sparse within ischemic myocardium at risk in Tau + I/ R group as compared with that in I/R group (TUNEL stain). Apoptosis rate in ischemic myocardium from I/R and Tau + I/R groups detected by flow cytometry was 17.66% +/- 1.54% and 4.86% +/- 1.23%, respectively. Fas and Bax protein expressions in ischemic myocardium of I/R group were higher than that in nonischemic myocardium group (P < 0.01), Bcl-2/Bax ratio in I/R group was lower than that in nonischemic myocardium (P < 0.01); while in Tau + I/R group, Fas and Bax protein expressions were lower than that in I/R group (P < 0.01), the Bcl-2/Bax ratio was higher than that in I/R group (P < 0.01).

CONCLUSIONTaurine reduced apoptosis of myocytes in I/R rabbit heart; its mechanism may involve Fas, Bax and Bcl-2 proteins expression.

Animals ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Male ; Myocardial Ischemia ; metabolism ; pathology ; Myocytes, Cardiac ; cytology ; drug effects ; Rabbits ; Reperfusion Injury ; metabolism ; pathology ; Taurine ; pharmacology