Angiogenesis Inhibitors ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Hepatocellular ; blood supply ; metabolism ; pathology ; therapy ; Chemoembolization, Therapeutic ; methods ; Endostatins ; therapeutic use ; Humans ; Liver Neoplasms ; blood supply ; metabolism ; pathology ; therapy ; Microvessels ; pathology ; Neovascularization, Pathologic ; pathology ; Vascular Endothelial Growth Factors ; metabolism

Background Hypertension is a multigenetic inheritable disease.Gene therapy with long-term effects and less side effects by regulating gene expression has been shown to be a potential and exciting prospect. Objective To investigate the effects of RNA interference(RNAi)targeting angiotensin-converting enzyme(ACE)on the blood pressure and ACE expression in kidney of spontaneously hypertensive rats(SHR).Methods SHR were randomly to receive placebo(n=12)or control adenovirus Ad5-EGFP)or a single injection of recombinant adenovi- ral vectors,Ad5-EGFP-ACE-shRNA(n=12,iv).Normotensive Wistar-Kyoto rats(WKY)were served as normal control group.SBP was measured before and after the intervention.Aorta,lung,myocardium and kidney were studied using fluorescence microscope to identify the sites of Ad5-EGFP-ACE-shRNA.Expressions of ACE mRNA and protein in kidney were evaluated by RT-PCR and Western blot.Results SBP of the treat group was effectively reduced by 19.0?3.2 mmHg at the 3rd day,and 22.1?3.3 mmHg at the 13th day of the experiment.The anti- hypertensive effect significant remained at least for 14 days.On the contrary,increase in BP was shown in placebo and the adenovirus control group.Compared with placebo or adenovirus control rats,ACE mRNA expression level in kidney of the treated rats was lower by 61.1% and 62.3% respectively,with ACE protein expression level lower- ing by 56.2% and 53.30% as well(ail P0.05). Conclusion RNA interference targeting ACE gene inhibits the expressions of ACE mRNA and protein.A single dose injection resulted in a prolonged decrease in BP.The evidence of strong antihypertensive effect by genetic therapy justifies efforts for further investigation.

OBJECTIVETo determine the ability of adhering and invading to host cells and the related pathologic changes of Leptospira spp. with different virulence.

METHODSA special Fontana silver staining method was developed to observe the ability of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601 and serogroup Pomona serovar pomona strain 56608, and L.biflexa serogroup Samaranga serovar patoc strain Patoc I to adhere Vero and J774A.1 cells. Ultrastructural lesions of the infected cells were examined by electron microscopy. By using flow cytometry with fluorescein labeling of FITC-Annexin V/PI,apoptosis and necrosis of the Vero and J774A.1 cells induced by the leptospiral strains before and after inactivation with ultraviolet treatment were detected, respectively.

RESULTSThe adhering rates of L.interrogans strains 56601 and 56608 to the Vero cells were 24.2% and 22.9% (P>0.05), while to the J774A.1 cells were 49.0% and 46.9% (P>0.05), respectively. L.biflexa strain Patoc I did not adhere to these two host cells. After the two strains of L.interrogans invaded two different cell lines, the special phagosomes containing the Leptospira were formed and similar cell ultrastructural lesions, such as chromatilysis and chromatin condensation, vacuolar degeneration, mitochondrion swelling and mitochondrial crista disappearance, and endoplasmic reticulum swelling and paramembranous ribosome disappearance, were observed. The apoptosis rates in the Vero cells caused by the L.interrogans strains 56601 and 56608 before and after ultraviolet inactivation were 84.4%, 82.8% and 77.9%, 86.1%, respectively. The L.interrogans strain 56601 mainly induced the terminal apoptosis in Vero cells with the rates of 68.0% and 52.9% before or after inactivation, while the L.interrogans strain 56608 mainly induced the early apoptosis in Vero cells with apoptosis rates of 64.1% and 50.1% before or after inactivation. In the J774A.1 cells, the L.interrogans strain 56608 caused cell necrosis (before and after inactivation) and apoptosis that was dominated by terminal apoptosis.

CONCLUSIONThe established Fontana silver staining method can be used to observe adhesion of L.interrogans.L.interrogans can invade host cells through endocytosis which causes untrastructural lesions. The necrosis or apoptosis induced by L.interrogans are affected by different host cell lines but not the virulence of L.interrogans strains.

Animals ; Apoptosis ; physiology ; Cell Adhesion ; Cells, Cultured ; Cercopithecus aethiops ; Endocytosis ; Humans ; Leptospira interrogans ; classification ; pathogenicity ; Macrophages ; microbiology ; ultrastructure ; Serotyping ; Vero Cells ; Virulence

OBJECTIVETo evaluate a retro-grade auricular flap for repairing large alar defects.

METHODSTwenty-nine adult cadavers were anatomically used for vascular investigation of the frontal, nasal and temporal regions by injecting a dye into the main vessels. Based on the anatomical study in these regions, a retro-grade auricular flap was designed for repairing alar defects in 16 patients.

RESULTSThe blood supply of the auricle could be nourished by the inner carotid artery system from the supratrachlear artery and supraorbital artery through the frontal arterial anastomotical network into the frontal branch of the superficial temporal artery. It is then passing the main trunk of the superficial temporal artery into the ear area through the auricular branches of the superficial temporal artery. The retro-grade auricular island flap could be formed by basing the supratrachlear artery and the supraorbital artery through the vascular network between the superficial temporal artery and the supratrachlear artery or the supraorbital artery. Sixteen patients with large alar defects and half-sized nasal defects were successfully repaired by this technique.

CONCLUSIONSThe retro-grade auricular island flap, based on the inner carotid artery system, could be a good and safe flap for repairing a large alar defect or half-sized nasal reconstruction.

Adult ; Arteries ; Cadaver ; Carotid Artery, Internal ; Ear Auricle ; blood supply ; Humans ; Nose Deformities, Acquired ; surgery ; Ophthalmic Artery ; Reconstructive Surgical Procedures ; Surgical Flaps ; blood supply ; transplantation ; Temporal Arteries

OBJECTIVETo compare botanical characteristics of cultivated Chrysanthemum morifolium cv. 'Hangju' of different origins in order to provide the basis for introduction and cultivation of Ch. morifolium cv. 'Hangju'.

METHODThe characteristics of plants, leaves and capitulum of Ch. morifolium cv. 'Hangju' were measured, and the obtained data were analyzed and compared.

RESULTThe range of plant height was 60.87-99.47 cm, number of branches 2.76-5.20, leave length 4.90-8.40 cm, leave width 3.25-5.38 cm, aspect ratio of leave 1.35-1.83, number of leave split 1.92-3.08. Numbers of capitulum were 21.92-53.12, diameter of capitulum 3.41-5.48 cm, lays of ray florets 3.28-7.16, number of ray florets 55.32-114.60, ray florets length 1.58-2.37 cm, ray florets width 0.50-0.69 cm, aspect ratio of ray florets 2.90-3.99, diameter of tubular flower 1.10-1.58 cm.

CONCLUSIONThe botanical characteristics of cultivated Ch. morifolium cv. 'Hangju' were distinguished from different origins. With the cultivation environment change, the botanical characteristics of the cultivars are changed.

China ; Chrysanthemum ; chemistry ; growth & development ; Flowers ; chemistry ; growth & development ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development

BACKGROUND AND OBJECTIVEThe level-Ib lymph node metastasis is rare in nasopharyngeal carcinoma (NPC). When and how this level should be irradiated with precise radiotherapy remains controversial. This study evaluated the prevalence and prognostic significance of level-Ib lymphadenopathy on the prognosis of NPC patients.

METHODSFrom January 1990 and December 1999, 933 newly diagnosed patients with NPC treated at Sun Yat-sen University Cancer Center were randomly selected, examined with computed tomography (CT) imagining for evidence of level-Ib lymphadenopathy before treatment. All patients received radical radiotherapy with or without chemotherapy. The relationship between level-Ib lymphadenopathy and post-treatment outcomes including overall survival (OS), locoregional recurrence-free survival (LRFS), and distant metastasis-free survival (DMFS) were analyzed using Kaplan-Meier methods. The Cox proportional hazards regression model was used to adjust for other prognostic factors.

RESULTSOf the 933 patients, 55 (5.9%) were found to have level-Ib lymphadenopathy, which was associated with carotid sheath involvement, oropharynx involvement and levels, and lateral cervical lymph node involvement. In the subgroup with carotid sheath involvement, with multivariate analysis accounting for all previously known prognostic factors, level-Ib lymphadenopathy was still associated with a risk of decreased OS (RR, 2.124; P<0.001), DMFS (RR, 2.168; P<0.001), and LRFS (RR, 1.989; P=0.001).

CONCLUSIONLevel-Ib lymphadenopathy in the patients with carotid sheath involvement is an independent prognostic factor.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; diagnostic imaging ; drug therapy ; pathology ; radiotherapy ; Chemotherapy, Adjuvant ; Child ; Cobalt Radioisotopes ; therapeutic use ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; radiotherapy ; Neck ; pathology ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Particle Accelerators ; Pharynx ; pathology ; Prognosis ; Proportional Hazards Models ; Radiography ; Radioisotope Teletherapy ; Retrospective Studies ; Survival Rate ; Young Adult

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .

Objective To evaluate the IL-33/ST2 signaling pathway′s role in the treatment of diastolic heart failure ( DHF) by using bisoprolol.Methods Twenty four diastolic heart failure rats were randomly divided into control group ( n=12 ) which were given distilled water and the biso-prolol group ( n=12) which were given solution of bisoprolol treatment.The treatment period lasted for 3 months.The data of left ventricular pressure , left ventricular filling time ( LVFT) , left ventricular end-dias-tolic pressure , the maximum rate of isovolumic left ventricular systolic pressure rise were observed in the two groups.The cardiomyocytes ST2 protein and expression of serum TNF -αwere compared.Results The left ventricular systolic pressure and left ventricular mean pressure were significantly lower in bisoprolol group than that in control group ( P <0.05 ) , but there was no statistical difference in left ventricular end -dias-tolic pressure ( P>0.05 ).The maximum rate pressure rise of left ven-tricular systolic content in bisoprolol group was significantly higher than that in the control group ( P<0.05 ) , and the maximum rate of isovolumic left ventricular diastolic pressure decreased significantly than that in con-trol group ( P <0.05 ) , but the LVFT was not statistically different between the two groups ( P<0.05 ).The expression of IL-33 protein in bisoprolol group was significantly lower ( P<0.05 ) , but the ST2 expression and serum TNF -αconcentration between the two groups was not statistically different ( P>0.05 ).Conclusion Bisoprolol may reduce left ventricular blood pressure in diastolic heart failure mice , and im-prove left ventricular compliance , which may be associated with a significant reduction in myocardial cells IL -33 pro-tein expression.

Objective： The current study was designed to investigate the effect of heat shock protein 70(HSP70) on stability of mutant p53 protein. Methods： Retroviral recombinant expressing antisense HSP70 RNA was constructed and transfected into MCF7/Adr breast cancer cells. The existence of foreign DNA was identified by PCR method and the HSP70 protein level was determined by Western blot analysis. The half-life of mutant p53 protein(mtp53) was measured by p53 stability assay. Results： The stable expressing strain(MAp70) from transfected cells was obtained through G418 selection. The foreign DNA in transfectant cells were confirmed by PCR, and the repression rate of HSP70 protein was 42％ . The half-life of mutant p53 in MAp70 cell was 12 hours, significantly lower than that of the control cells. Conclusion： Antisense HSP70 RNA can decrease the HSP70 protein level and significantly increase instability of mutant p53 protein in MCF7/Adr breast cancer cell.

Objective To investigate the effects of RNA interference(RNAi)targeting angiotensinconverting enzynle(ACE)on the blood pressure and myocardial remodeling in spontaneously hypertensive rats(SHRs).Methods Saline(control),adenovirus(Ad5)and recombinant adenoviral vectors(Ad5-ACE-shRNA expressing ACE gene-specific shRN)were randomly administered by caudal intravenation to SHRs(n=12 each group)at day 1 and day 16.Normotensive Wistar-Kyoto rats(WKY)served as normal controls.Systolic blood pressure(SBP)of the caudal artery was measured daily.Expressions of ACE at mRNA and protein levels in myocardium and aorta were evaluated by RT-PCR and Western blot respectively,ACE serum concentration was measured by ELISA at day 3(n=6 each group).The ratio of left ventricular to body weight(LVW/BW),myocardial collagen content were measured and myocardial uhrastrueture observed under transmission electron microscope at the study end.Results Ad5-ACE-shRNA injection significantly reduced SBP(-22 mm Hg,1 mm Hg=0.133 kPa)and the antihypertensive effect could last at least 14 davs post each injection.SBP was not affected by saline and Ad5 injections.ACE expressions at mRNA and Drotein levels at myocardium and aorta as well as serum ACE were significantly decreased in Ad5-ACE-shRNA treated SHRs compared to that in saline and Ad5 groups(all P＜0.05)and was coinparable to that in WKY group(P＞0.05).The LVW/BW ratio(2.24±0.19)and myocardial collagen content[(1.283±0.019)μg/mg]in Ad5-ACE-shRNA treated SHRs were also significantly lower than those in saline treated[3.21±0.13 and (1.686±0.013)μg/mg,both P＜0.05]and Ad5 treated SHRs [3.13 ±0.12,(1.682±0.009)μg/mg,both P＜0.05]but still higher than those of WKY group[2.06±0.11,(1.257±0.019)μg/mg,both P＜0.05].Myocardial uhrastructure was also significantly improved in Ad5-ACE-shRNA treated SHRs compared to saline and Ad5 treated SHRs.Conclusion RNAi targeting ACE gene significantly inhibited the expressions of ACE at mRNA and protein levels and resuhed in prolonged antihypertensive effects and myocardial uhrastructure improvements in this SHR model.