OBJECTIVE:To investigate Smad3 and Smad7 expression in cardiac tissues in myocardial infarction (MI) rats and the effect of simvastatin (SIM) on expression of Smads.METHODS:36 rats were randomized into SIM (40 mg?kg-1,n=12) group,MI group (normal saline,n=12) and SHAM group (normal saline,n=12).Rats in first two groups were induced MI model.Rats of 3 groups were sacrificed after four weeks administration.The mRNA expressions of Smad3 and Smad7 in non-infarction regions were measured by RT-PCR.Smad3 and Smad7 protein expressions were measured by Western blotting assay.RESULTS:As compared with SHAM group,the expressions of Smad3 in MI group and SIM group were increased while the expressions of Smad7 were decreased.As compared with MI group,the mRNA expression and protein expression of Smad3 in SIM group was decreased while that of Smad7 increased (P

BACKGROUND: A fibrinolytic enzyme (FLE) was purified from the venom of Agkistrodon blomhoffi brevicaudus by ion-exchange chromatography;meanwhile, it can dissolve thrombotic anti-thrombus drugs during onset of myocardial infarction and apoplexy.OBJECTIVE: To investigate the purification of FLE from the venom of Agkistrodon blomhoffi brevicaudus and research some biochemical and pharmacological characterizations.DESIGN: Controlled observational study.SETTING: Snake Venom Research Institute, Guangxi Medical University.MATERIALS: The experiment was carried out in Snake Venom Research Institute of Guangxi Medical University from July 2003 to June 2005.Kunming mice, weighing 18-25 g, of mean body mass of (20±2) g, aged 3-4 months were provided by Experimental Animal Center of Guangxi Medical University. Agkistrodon blomhoffi brevicaudus was supplied by Snake Venom Research Institute of Guangxi Medical University.METHODS: FLE was purified with DEAE Sepharose CL-6B ion exchang columns and HiPrep Sephacryl S 100; purity and relative molecular mass were measured with high-effective chromatograph of liquid and polyacrylamidedel electrophoresis (PAGE); isoelectric point was measured with isoelectric focusing electrophoresis; and then, components and partially pharmacological characteristics were also measured.MAIN OUTCOME MEASURES: Relative molecular mass, conformation of isoelectric point and amino acid, effect of inhibitor on FLE, hemorrhagic activity, and anticoagulant effect of FLE.RESULTS: All 30 rats and 12 rabbits were involved in the final analysis.Relative molecular mass was 59 100 and isoelectric point was 4.98. SDSPAGE acted as a zone in vitro. High-effective chromatograph of liquid showed a simple peak of FLE. This group belonged to metallo proteinases.Analysis of components of amino acid suggested that it contained a lot of acidity amino acids, had mild hemorrhagic toxicity and strongly thrombolytic effect.CONCLUSION: A simple acidity metallo proteinases, which is characterized by mild hemorrhagic toxicity and strong anti-coagulation, can be purified from the venom of Agkistrodon blomhoffi brevicaudus.

Objective Atrioventricular block (AVB) is a common and serious arrhythmia. At present, there is no perfect method of treatment for this kind of arrhythmia. The purpose of this study was to regenerate cardiac atrioventricular conduction by autologous transplantation of bone marrow mesenchymal stem cells (MSCs), and explore new methods for therapy of atrioventricular block. Methods Eleven Mongrel canines were randomized to MSCs transplantation (n=6) or control (n=5) group. The models of permanent and complete AVB in 11 canines were established by ablating His bundle with radiofrequency technique. At 4 weeks after AVB, bone marrow was aspirated from the iliac crest. MSCs were isolated and culture-expanded by means of gradient centrifugal and adherence to growth technique, and differentiated by 5-azacytidine in vitro. Differentiated MSCs (1ml, 1.5×107cells) labeled with BrdU were autotransplanted into His bundle area of canines by direct injection in the experimental group, and 1ml DMEM in the control group. At 1-12 weeks after operation,the effects of autologous MSCs transplantation on AVB models were evaluated by electrocardiogram, pathologic and immunohistochemical staining technique. Results Compared with the control group, there was a distinct improvement in atrioventricular conduction function in the experimental group. MSCs transplanted in His bundle were differentiated into analogous conduction system cells and endothelial cells in vivo, and established gap junction with host cardiomyocytes. Conclusions The committed-induced MSCs transplanted into His bundle area could differentiate into analogous conduction system cells and improve His conduction function in canine AVB models.

Objective To explore the feasibility of using composite scaffolds of rabbit oral epithelial cells and polycaprolactone (PCL) electrospun fibers for urethral repair. Methods The 25%PCL was prepared using a 5:1 by volume mixture of trichloromethane and anhydrous methanol, and PCL fiber tubular scaffolds were obtained by electrospinning. Rabbit oral mucosa epithelial cells (1.5 × 105) were implanted on the PCL scaffold. Subsequently, they were embedded in nude mice subcutaneous, explanted in 2 weeks. PCL fiber tubular scaffolds without rabbit oral mucosa epithelial cells were used as control. The complex urethral scaffolds were evaluated by immunofluorescence staining with cytokeratin antibody and HE staining. Results Compared with blank PCL group, the rabbit oral mucosa epithelial cell group showed a good cellularization. Rabbit oral mucosa epithelial cells formed a dense cell layer on the surface of PCL lumen, which suggested that rabbit oral mucosa epithelial cells can proliferate on the surface of PCL lumen. Conclusion Rabbit oral epithelial cells can be used as one of the seed cells for tissue engineered urethral scaffolds, and it is possible to construct tissue engineering substitute materials for urethral repair by rabbit oral epithelial cells combined with PCL.

OBJECTIVE:To compare in vivo and in vitro permeation behaviors of the ethosome gels of testosterone,testoster-one propionate and testosterone undecanoate. METHODS:The ethosome gels of testosterone,testosterone propionate and testoster-one undecanoate were prepared. With cumulative permeating amount and permeation rate as the indexes,Franz diffusion cell and HPLC were employed to compare in vitro permeation behaviors of 3 kinds of ethosome gels in mouse skin. With testosterone patch as the positive control drug, electrochemistry method was adopted to detect the concentration of testosterone in plasma 0,3,6, 9,12,24,36 and 48 h after applying such 3 kinds of ethosome gels on the back of rats,and then pharmacokinetic parameters were calculated with DAS 2.0 software. RESULTS:24 h cumulative permeating amounts of the ethosome gels of testosterone,tes-tosterone propionate and testosterone undecanoate were(234.31±13.8),(175.63±41.1)and(72.60±15.3)μg/cm2,and the per-meation rates were(10.25±1.9),(7.64±1.4)and(2.96±0.8)μg/(cm2·h),respectively. The pharmacokinetic parameters of the above-mentioned three kinds of ethosome gels and the positive control drug were respectively as follows as cmax of(20.19±2.57), (17.50±2.91),(0.23±0.04),(14.97±2.12)ng/ml,t1/2Ka of(2.80±0.45),(3.36±0.59),(4.02±0.62),(4.20±0.71)h,AUC0-48 h of(13.85±1.96),(13.93±2.13),(0.35±0.07),(11.76±2.31)ng·h/ml. CONCLUSIONS:in vivo and in vitro permeation behav-iors of the ethosome gels of testosterone and testosterone propionate are fairly good.

A simple and efficient method for fabricating a novel surface-enhanced Raman scattering ( SERS) substrate with good reproducibility and high SERS activity was reported. Cu2 O was prepared by mixing CuCl2 ·2H2 O with ascorbic acid, which was then used as the templates for depositing of gold nanoparticles ( AuNPs) on their surfaces, forming Cu2 O@ Au with heterostructures. Transmission electron microscopy, scanning electron microscopy and X-ray diffraction observation revealed that Cu2 O had polyhedral structure and smooth surface, and AuNPs were closely deposited on the surface of Cu2 O. It was used as SERS substrate for detection of Rhodamine B with linear detection range of 1 × 10-2-5 × 10-6 mol/L, and detection limit of 3 ×10-7 mol/L. Cu2 O@Au showed good chemical stability, remained stable in acid, PBS and river samples, and could be used in the SERS detection of target in water sample.

Animals ; Brain Edema ; Brain Ischemia ; Cerebral Infarction ; Cerebrovascular Circulation ; Rats ; Reperfusion ; Reperfusion Injury

To study the immunopathological characteristics and differential diagnosis of hepatic angiomyolipoma(AML).Methods:Thirty-six surgically resected hepatic AML were investigated clinicopathologically and immunohistochemically with 10 antibodies.Results:Hepatic AML occurred in 21 females and 15 males,with the mean age of 41.6 years(26-60 years old).The patients with AML often had no special symptoms even had large space-occupying lesions in the liver.The diameter of AML was 2.5 cm to 14 cm(mean 6.8 cm).Histologically,AML was composed of varying heterogeneous mixture of 3 tissue components:blood vessels,smooth muscle and adipose cells.Extramedullary hemopoiesis sometimes existed.According to tissue components,AML was subcategorized into mixed type(19.4%,n=7),lipomatous type(11.1%,n=4),myomatous type(66.7%,n=24),and angiomatous type(2.8%,n=1).The epithelioid smooth muscle cells were sensitive to HMB-45(100%),SMA(100%),and CD117(66.7%) staining.Conclusion:Hepatic AML often contains smooth muscle elements,which have varied morphological features and should be carefully differentiated from hepatocellular carcinoma,mesenchymal hamartoma，and tumors with rich fat or blood vessels.Immunohistochemical staining with HMB-45 and SMA are the best available markers for the diagnosis of hepatic AML.

Natural killer (NK) cells, as an essential part of innate immunity, can directly identify and kill tumor cells after being activated by the synergistic action of surface inhibitory receptors and activated receptors. It can secrete cytokines to recruit dendritic cells (DCs), induce DCs maturation and enhance adaptive immune response. It can target cancer stem cells (CSCs) and circulating tumor cells (CTCs) to inhibit cancer metastasis. NK cells have a unique inflammatory tendency, which can respond to cytokines and chemokines released from tumor sites and migrate to tumor sites, making them occupy an important advantage in cancer targeted therapy. The research on cancer targeted therapy of NK cells as drug delivery carriers, NK cell membrane-coated biomimetic nanoparticles, and NK cell extracellular vesicles (NKEVs) has attracted more and more attention. The article will focus on the mechanism of NK cells inhibiting cancer, and summarize the research progress of cancer targeted therapy of NK cells.

Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P