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 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 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 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

The brain pharmacokinetics of hyperoside and 1 , 5-dicaffeoylquinic acid and the effects of acanthopanax senticosus leaves on neurotransmitters in cerebral ischemic rat brain were investigated. In the study, acute incompleteness cerebral ischemia model was developed by ligating the bilateral common carotid arteries of rats. The differences of brain pharmacokinetics of hyperoside and 1,5-dicaffeoylquinic acid between control and cerebral ischemic rats were determined by online microdialysis coupled with MS/MS method. At the same time, the contents of glutamic acid (Glu), aspartic acid (Asp), γ-aminobutyric acid (GABA), serotonin (5-HT) , dopamine ( DA) , and acetylcholine( Ach) in rat hippocampus among different groups were determined simultaneously in MRM mode. ACE 5 C18-AR column was used for separation. Results showed that the online analytical method had excellent linearity (R2>0. 99). The accuracy and precision could meet the analysis requirement. Besides, the hyperoside and 1,5-dicaffeoylquinic acid penetrated the blood-brain barrier successfully and metabolized quickly, but the absorption reduced and elimination became faster in cerebral ischemic rats; the contents of Glu, Asp, GABA and DA in brain issue of cerebral ischemic rats decreased significantly ( p<0 . 01 ) and the level of Ach increased remarkably ( p<0 . 05 ) after a preventive medication with the extract of acanthopanax senticosus leaves for one week.

Objective To investigate the level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cell in peripheral blood and hydro-thorax of the TB patients and its clinical significance .Methods 49 patients with tuberculosis(TB) including 14 cases of tuberculous pleurisy and 27 individuals with latent TB infection were selected and 66 healthy individuals were selected as the controls .PMA and ionomycin were adopted to stimulate mononuclear cells in whole blood and pleural effusion .The secretion status of CD4+ T cells cy-tokines was detected by using the intracellular cytokine staining and the flow cytometric analysis .Results According to the differ-ent cytokines generated by CD4+ T cells ,which were divided into 7 cell subgroups :IL-2+ IFN-γ+ TNF-α+ ,IL-2+ IFN-γ+ ,IL-2+TNF-α+ ,IFN-γ+ TNF-α+ ,IL-2+ ,TNF-α+ and IFN-γ+ cell subgroups .The proportion of peripheral blood IL-2+ IFN-γ+ TNF-α+multifunctional Th1 cells in the TB patients was significantly lower than that in the healthy controls and the individuals with latent TB infection(P<0 .01) ,the expression levels of IL-2+ IFN-γ+ cells and IFN-γ+ TNF-α+ cells were significantly lower than those in the individuals with latent TB infection and the healthy controls (P<0 .05);TNF-α+ cells was higher than that in the healthy con-trols and the individuals with latent TB infection (P<0 .05) .The other subgroups had no obvious change .The response level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cells in the pleural effusion mononuclear cells (PEMC) was higher than that in the peripher-al blood mononuclear cells(P<0 .05);IL-2+ cells in peripheral blood mononuclear cells (PBMC) was lower than that in PEMC (P<0 .01) .Conclusion The response of non-specific Th1 cells is related with the clinical outcome of TB infection ,IL-2+ IFN-γ+TNF-α+ multifunctional Th1 cells plays a certain role in the protective immunoreaction of TB .

OBJECTIVETo investigate the clinicopathological features of testicular malignant Leydig cell tumor (TMLCT) and improve the non-invasive diagnosis of the disease.

METHODSWe retrospectively analyzed the clinicopathological data on a case of TMLCT, detected the circulating tumor cells (CTC) in the peripheral venous blood, and reviewed the related literature.

RESULTSThe patient, a 47-year-old male, underwent radical orchidoepididymectomy under general anesthesia. Postoperative pathology confirmed the lesion to be TMLCT, which was mainly composed of Leydig cells and suspected with vessel carcinoma embolus. Immunohistochemistry showed the tumor cells to be positive for α-inhibin, Ki67, CD30, vimentin, EMA, and PLAP, but negative for CK, CK7, S100, CD10, SMA, Des, AFP, hCG, CEA, CK19, CD117, Oct-4, LCA, CD20, Pax-5, CD3, and CD43. Two CTCs were detected in the peripheral venous blood. The patient received 3 courses of chemotherapy for retroperitoneal multiple lymph nodes metastasis post-operatively. Subsequent CT imaging manifested no obvious reduction of the retroperitoneal lymph nodes and consequently the patient again underwent retroperitoneal lymphadenectomy and cryoablation. At 8 months after treatment, CT examination revealed notably enlarged retroperitoneal lymph nodes with the right adrenal gland evidently invaded.

CONCLUSIONTMLCT is an extremely rare sex-gonad stromal tumor with high malignancy and poor prognosis, and CTCs may be used for its early diagnosis and prognostic prediction.

Biomarkers, Tumor ; metabolism ; Humans ; Immunohistochemistry ; Leydig Cell Tumor ; drug therapy ; pathology ; surgery ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; Prognosis ; Retrospective Studies ; Sex Cord-Gonadal Stromal Tumors ; drug therapy ; pathology ; surgery ; Testicular Neoplasms ; drug therapy ; pathology ; surgery