OBJECTIVETo evaluate the impact of nicotine- and tar-free cigarette smoke extract (fCSE) on the serum testosterone (T) level and erectile function of male rats.

METHODSWe randomized 30 male SD rats to three groups of equal number to receive subcutaneous injection of PBS (1.0 ml / 300 g body weight per day), fCSE (1.0 ml/300 g body weight per day), and reduced glutathione hormone (GSH, 200 mg per kg body weight per day) in addition to fCSE (fCSE + GSH), respectively, all for 8 weeks. Then we evaluated the erectile function of the rats by measuring the maximal intracavernous pressure (MICP), mean arterial pressure (MAP), ICP/MAP ratio, time of stimulation to MICP (Tmax), and cavernosal filling fate (CFR). We determined the serum T level, the activities of superoxide dismutase (SOD) , malondialdehyde (MDA), and nitric oxide synthase (NOS) in the cavernosal tissue, and also observed the morphological changes of the corpus cavernosum.

RESULTSCompared with the controls, the rats of the fCSE group showed obvious decreases in the levels of serum T ([5.37 ± 1.43] vs [3.22 ± 1.11] μg/L), NOS ([2.90 ± 0.27] vs [1.67 ± 0.18] U/mg) , and SOD ([18.41 ± 1.09] vs [13.36 ± 1.18] U/mg prot) and erectile function-related indexes MICP ([85.92 ± 6.36] vs [58.99 ± 10.76] mmHg), MICP/MAP (0.86 ± 0.09 vs [0.56 ± 0.08]), and CFR (2.14 ± 0.44 vs 0.89 ± 0.44), but markedly increased Tmax ([29.90 ± 5.78] vs [42.90 ± 8.56]s), with a positive correlation between the serum T level and CFR (r = 0. 364, P < 0.05). Masson staining revealed a lower ratio of the corpus cavernosum smooth muscle tissue to collagen fiber in the fCSE group (0.27 ± 0.04) than in the control (0.98 ± 0.12). Compared with the fCSE group, the fCSE + GSH group exhibited significantly improved MICP ([58.99 ± 10.76 ] vs [77.95 ± 7.71] mmHg), MICP/MAP (0.56 ± 0.08 vs 0.77 ± 0.09), and CFR (0.89 ± 0.44] vs 1.76 ± 0.42) and shortened Tmax ([42.90 ± 8.56 ] vs [32.10 ± 5.84 ] s). The ratio of the corpus cavernosum smooth muscle tissue to collagen fiber was higher in the fCSE + GSH than in the fCSE group (0.77 ± 0.09 vs 0.27 ± 0.04) but still lower than in the control (0.98 ± 0.12).

CONCLUSIONNicotine- and tar-free cigarette smoke extract reduces the serum T level and erectile function of rats, which is related to oxidative stress. Antioxidant therapy can improve erectile function but has a limited value for morphological protection of the penile tissue.

Animals ; Erectile Dysfunction ; chemically induced ; Male ; Malondialdehyde ; metabolism ; Muscle, Smooth ; pathology ; Nicotine ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Superoxide Dismutase ; metabolism ; Tars ; Tobacco ; adverse effects

We aimed to study the effect of allitridum (All) on the transient outward potassium current (Ito) of ventricular myocytes of spontaneously hypertensive rats (SHR). Totally 30 male SHRs were randomly divided into three groups: low-dose All group (7.5 mg·kg(-1)), high-dose All group (15.0 mg·kg(-1)) and normal saline group. The other 10 sex and age matched Wistar-kyoto rats (WKY) were also taken as control group (WKY group). All animals received i.p. administration for 8 weeks. The dual enzymatic method was used to separate single ventricular myocyte from animals. Patch-clamp technique was used to record Ito and analyze the effect of All on the current. It was shown that the left ventricular hypertrophy of SHR was reversed significantly by All. Furthermore, the density of Ito was recovered in both high and low dose All groups. The peak current densities of Ito were enhanced from 18.23±3.64 to 25.17±2.86 pA/pF (P<0.01) and 36.47±5.42 pA/pF (P<0.01) at +50 mV by All 7.5 mg·kg(-1) and 15.0 mg·kg(-1), respectively, which was not significantly different with WKY group. The effect was associated with positive shift of the steady-state, close-state inactivation, and shortened recovery from inactivation of Ito. It is concluded that All decreases the remodeling of Ito of ventricular hypertrophic myocytes of SHR.
Allyl Compounds ; pharmacology ; Animals ; Hypertrophy, Left Ventricular ; drug therapy ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Sulfides ; pharmacology

Objective To investigate the factors associated with delay in decision to seek treatment in patients with acute myocardial infarction(AMI) in Beijing. Methods This prospective,cross-sectional,multicenter survey was conducted from November 1,2005 and December 31 ,2006. The participants consisted of 799 patients with STEMI admitted within 24 h of symptom onset to 19 hospitals in Beijing. Data were collected by semi-structured interviews and medical records review. The patients were categorized into an early decision group and the a late decision group based on the 30 min cut-off. Results The median(25%,75%) decision delay in STEMI patients was 60(20, 180)min. Factors associated with late decision in an univariate analysis were age ≥65 years, retirement or unemployment, history of myocardial infarction,symptom onset at home and intermittent symptoms, whereas presence of bystanders such as friends,coworkers or even strangers,unbearable symptoms,dyspnea,sweating,syncope and attribution of symptoms to cardiac origin were related to early decision. Multivariate logistic analysis showed that history of myocardial infarction,absence of syncope, intermittent symptoms,bearable symptoms and attribution of symptoms to noncardiac origin were independent predictors of decision delay＞30 min. Patients in the early decision group had more chances to receive acute reperfusion therapies(P=0.001) and shorter time intervals from symptom onset to reperfusion therapies(P＜0.001). Conclusions To a great extent patients with AMI in Beijing delayed in decision to seek treatment. History of myocardial infarction, symptom characteristics and symptom attribution were associated with decision delay.

OBJECTIVEThis study aims to detect microRNA-17(mir-17) expression on the osteogenic differentiation of advanced glycation end products (AGEs)-stimulated hunman periodontal ligament stem cells (HPDLSCs) and to analyze the influence of these cells on this process.

METHODSHPDLSCs were isolated using limited dilution technique. After osteogenic differentiation occurred, different time points of mir-17 expression in the experimental groups were detected by real time polymerase chain reaction (PCR). The mir-17 overexpression and inhibition were evaluated using cell transfection technique. Differences in gene expressions were detected by real time PCR; differences in protein expressions were analyzed by Western blot.

RESULTSThe mir-17 expression was reduced after osteogenic differentiation occurred at 3, 7, and 14 d compared with that in the control group (P < 0.05). The expression levels of bone sialoprotein (BSP), Runt-related transcription factor-2 (Runx-2)and alkaline phosphatase (ALP) in the experimental groups were lower than those in the mimic control group when mir-17 expression increased. In addition, the protein expression levels of Runx-2 in the experimental groups were lower than those in the control group. The expression levels of BSP, Runx-2 and ALP in the experimental groups were higher than those in the inhibitor control group when mir-17 expression decreased. Likewise, the protein expression levels of Runx-2 in the experimental groups were higher than those in the control group.

CONCLUSIONAGEs inhibit the osteogenic differentiation of HPDLSCs by affecting mir-17 expression.

Alkaline Phosphatase ; Cell Differentiation ; Glycation End Products, Advanced ; Humans ; MicroRNAs ; Osteogenesis ; Periodontal Ligament ; Stem Cells

OBJECTIVEThe effect of advanced glycation end products (AGEs) on the osteogenic differentiation of humanperiodontal ligament stem cells(hPDLSCs) was discussed. Changes in the Wnt signaling pathway during glycation were also determined.

METHODSIn vitro tissue explanting method was primarily applied. Limiting diluted clone was cultured to obtain hPDLSCs in vitro. The subjects were divided into two groups: the healthy group (N-hPDLSCs) and the AGEs-stimulating group (A-hPDLSCs). Osteoblast mineralization was induced in the experimental groups. The following processes were performed: alizarin red staining; alkaline phosphatase (ALP) staining; real time polymerase chain reaction (real time PCR) for detecting osteogenic genes and Wnt classical pathway-related factors, DKK-1 and β-catenin; Western blot analysis. Bone protein and β-catenin were correlated in the nuclear expression.

RESULTSThe cells were osteogenically induced. ALP staining showed that the N-hPDLSCs displayed the deepest color. Alizarin red staining indicated that the A-hPDLSCs group had less calcified nodules than the N-hPDLSCs group. The real time PCR results suggested that the expression of relative osteogenic genes in A-hPDLSCs was quite low. Statistically significant differences in differentiation were found between groups (P < 0.05). The Western blot result was similar to that of real time PCR. Classical Wnt signaling pathway-related factor β-catenin was higher in A-hPDLSCs than in N-hPDLSCs. By contrast, DKK-1, which is an inhibitor in the Wnt pathway, had a significantly lower expression rate in A-hPDLSCs than in N-hPDLSCs. The Western blot result also showed that β-catenin expression in the nucleoprotein in A-hPDLSCs was notably higher than in N-hPDLSCs.

CONCLUSIONAGEs can inhibit hPDLSCs osteogenic differentiation. AGEs induce changes in the normal periodontal ligament stem cells classical Wnt pathway. Canonical Wnt pathway is reactivated because of AGEs stimulation.

Cell Differentiation ; Glycation End Products, Advanced ; Humans ; In Vitro Techniques ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Stem Cells ; Wnt Proteins ; Wnt Signaling Pathway ; beta Catenin

Objective To build the experimental basement for the clinical use of cytokines induced killer(CIK)cells from the cord blood mononuclear cells(CBMNC)in tumor adoptive cellular immunotherapy, an effective protocol for their proliferation in vitro and cytotoxicity of CIK cells was established.Methods The lymphocytes from umbilical cord blood were isolated by density gradient centrifugation and suspended in medium with CD_3 mAb,rIL-2,rIL-1 and IFN-? as inducing agents to prepare CIK cells.At the same time, the lymphokine activated killer(LAK)and CBMNC were set as controls,which were only added IL-2 and not any cytokines during the whole culture.The changes of CIK cells before and after induction were observed with microscope and the phenotypes of the cells were analyzed by using flow cytometry.The proliferation of CIK cells were determined by trypan blue exclusion assay and the cytotoxic activity to lung cancer cell were tested with MTF method.Results According to the experiment,combining use of four types of cytokines could generate a great deal of CIK cells possessing highly cytotoxicity.From day 5 CIK cells became to prolif- erate and reached the peak at day 14.During the whole period,the relative percentage of CD_3~+ CD_(56)~+ cells in- creased significantly.Compared with LAK cells,which reached the proliferation peak at day 7 and then showed no evident proliferation.The control cells(CBMNC)showed no evident change of phenotypes and proliferation.CIK cells showed a higher antitumor activity on the tumor cells than LAK cells and CBMNC in vitro.Conclusion Umbilical cord blood can generate a great deal of CIK cells combining used with cy- tokines.Compared with classic LAK cells,umbilical cord blood CIK cells have the advantages of rapid prolif- eration speed and powerful cytotoxicity.CIK cells will be promising as a new strategy for the adoptive cellular immunotherapy of tumor.

BACKGROUND:The measure and prediction of the shape of optic nerve head are the key issues for early diagnosis and duration prediction of glaucoma. Therefore, it is significant to research the shape variation of optic nerve head under high intraocular pressure.
OBJECTIVE:To establish three-dimensional finite element model of optic nerve head and to analyze the deformation of optic nerve head and the change in retina thickness after acute high intraocular pressure.
METHODS:Animal model of acute ocular hypertension was established by methods of anterior chamber perfusion. The tomographic images of the optic nerve head of a cat were obtained at the normal intraocular pressure and 5 320 Pa, 7 980 Pa, 10 640 Pa, 13 300 Pa, 15 960 Pa intraocular pressures using optical coherence tomography. Then, we measured the variation of retinal thickness at typical location. Basing on the tomographic images of optic nerve head of a cat at normal intraocular pressure, we obtained three-dimensional structure of retina and choroid using software MIMICS. Then three-dimensional model of optic nerve head was established by assembling the retina and choroid. The deformation of optic nerve head and the change in retinal thickness under high intraocular pressure were observed using software ABAQUS. The effectiveness of the model was verified by the experimental result.
RESULTS AND CONCLUSION:With the increase of intraocular pressure, the retinal thickness is thinner, the optic nerve head depth, width and the ratio of the depth to width are gradual y increased. It suggests that the acute high intraocular pressure causes retinal thinning, optic nerve head widening and deepening. It is feasible to establish optic nerve head modeling in vivo by using optical coherence tomography, mechanical analysis can be applied to predict the shape variation of optic nerve head, which is significant to further deduce the pathological process of glaucoma.

OBJECTIVE To investigate the uses of antibacterials,the characteristics of bacterial distribution and the drug resistance and provide a foundation for reasonable application of antibacterials.METHODS Germs isolation and cultivation were carried out according to National Clinical Testing Operation Procedures(2nd edition).Germs definition and resistance test were carried out by using VITEK 2 Compact microbiological assay system.Statistical analysis was processed by using WHONET 5.4.RESULTS Totally 7815 strains of bacteria were isolated from clinical samples,most of which were Gram-negatives(G-)(58.0%,56.3% and 59.5%,respectively in the 3 years).The tendency of Gram-positives(G+) didnot charee obviously(about 20%).Theresistance were increasing.The resistance rate of Staphylococcus aureus was more than 70.0 % to the most antibacterials.Up to now,there was no vancomycin-resistant isolate.CONCLUSIONS We should use antibacterials reasonably and efficiently to delay and control drug resistance,according to the susceptibility test.

AIM: To study the current density of transient outward potassium current (I_(to)) in cells from the epicardial zone of the 1-week and 2-month infarcted rabbit heart. METHODS: Rabbits were infarcted by ligation of the left anterior descending coronary artery, 1 week as well as 2 months later, the single ventricular myocytes were isolated enzymatically from the infracted area of 1-week infracted rabbit heart (PMI-1 week) and 2-month infracted heart (PMI-2 months), region remote from the infracted zone of 2-month infracted heart (REM-2 months) and free wall of left ventricule from noninfarcted heart (CON). I_(to) was recorded using whole cell patch-clamp techniques. (RESULTS:) Membrane capacitance of myocytes in REM-2 months group was signifitantly larger than that in CON. I_(to)current density (at +60 mV) was significantly reduced in PMI-1 week [(7.5?2.4) pA/pF, n=12] and PMI-2 months [(10.6?4.1) pA/pF, n=18] compared with CON [(17.4?5.2) pA/pF, n=16], P

AIM: To study the effect of experimental acute necrotizing pancreatitis (ANP) on sodium and L-type calcium current in rat cardiomyocytes. METHODS: I Na and I Ca-L were recorded using whole cell patch-clamp techniques from left ventricular myocytes in ANP model established by retrograde injection of 3 5% sodium taurocholate 2 5 mL/kg into pancreatic duct. RESULTS: Peak I Na current density (at -30 mV) was significantly reduced in ANP [(12 45?2 26) pA/pF, n =16] compared with sham [(25 32?3 31) pA/pF, n= 14], P