BACKGROUND:Transforming growth factor beta-1(TGF-β1)is a cytokine having variously biological effects in repair and renew of tissue injuries; meanwhile, tendon sheath fibroblasts and collagen Ⅰ play important roles in healing and desmoplasia of tendon.OBJECIVE:To study the effects of TGF-β1 on the proliferation and collagen production of tendon sheath fibroblasts.epitenon tenocytes and endotenon tenocytes in the three cell types of rabbit fexor tendon.DESIGN:Contrast observation study.SETTING:Department of Trauma Surgery,Affiliated Hospital of Medical College,Qingdao University.MATERIALS:The experiment was carried out in the Animal Laboratory,Affiliated Hospital of Medical College,Qingdao University from July 2004 to September 2005.A total of 6 adult New Zealand rabbits,of either gender,weighing 3.5-4.5 kg,were selected from Qingdao Experimental Animal Center.Collagenase was provided by Sigma Company;collagen Ⅰ,Ⅱand Ⅲ antibody by Sigma Company;TGF-β1 by Wuhan Boster Biology Company.METHODS: Three cell lines of tendon sheath,epitenon and endotenon were isolated from rabbit flexor tendon and cultured in serum culture media and then in serum-free culture media.In addition,the cells in the experimental group were added with 5 μg/L TGF-β1 in each well,but they were not added with any additive in the control group.MAIN OUTCOME MEASURES:①Proliferation in the two groups was measured with cytometry at 1,2,3 and 4 days after culture.②Preduction of collagens Ⅰ,Ⅱ and Ⅲ was measured with immunohistochemical staining at 4 days after culture.③Collagen contents of the three types were measured with enzyme linked immunosorbent assay(ELISA)in the two groups;expressJon of collagen Ⅰ gene was detected with reverse transcription polymerase chain reaction(RT-PCR).④Contents of collagen Ⅰ induced by TGF-β1 in various dosages of 0,5.10,15 and 20 μg/L were detected with ELISA technique.RESULTS:①Proliferated rates were similar in the two groups at 1 day after culture;however,proliferated rate of tendon sheath fibroblasts was rapidly increased, and there was significant difference as compared with that of epitenontenocytes and endotenon tenocytes(P＜0.05).②Expressions of collagens Ⅰ, Ⅱ and Ⅲ:Immunocytochemical stain demonstrated that three kinds of cells could produce collagens Ⅰ, Ⅱ and Ⅲ;while ELISA indicated that the contents of collagens in three types produced by tendon sheath fibroblasts were the most;in addition,content of collage Ⅰ was higher in the experimental group than that in the control group(P＜0.05-0.01).③Expression of collage Ⅰ gene of tendon sheath fibroblasts was increased as 1.3 times in the experimental group as that in the control group and there was signiflcant difierence(P＜0.01);meanwhile,expressions in epitenon tenocytes and endotenon tenocytes were also higher in the experimental group than those in the control group(P＜0.05).④TGF-β1 in the dosage of 5-10 μg/L had obvious effects on increasing production of collagen;however,production of collagen was not obviously changed when it was affeCted by TGF-β1 in the dosage of 10-20 μg/L.CONCLUSION: TGF-β1 can increase the production of collagen in tendon sheath fibroblasts,epitenon tenocytes and endotenon tenocytes and the expression of collagen Ⅰ gene. In addition, it is important for regulating level of TGF-β1 after tendon injury to prevent adhesion of tendon.

PurposeWith the extensive use of percutaneous radiofrequency ablation (RFA) for the treatment of hepatic carcinoma (HC), the study of MRI findings and its clinical signiifcance after RFA of HC have important value and can improve the complete ablation rate.Materials and MethodsA retrospective analysis of post-procedure MRI ifndings of 79 patients (114 lesions) with HC were performed, the size of the lesion, the signal changes and enhancement condition were observed at the ifrst, fourth and seventh month after RFA; the two different ifndings of high signal ring on MRI T1WI and local recurrence rate were analyzed.ResultsOne month after RFA, peripheral region of RFA lesion showed high signal on T1WI, and slightly lower signal on T2WI, the size of lesions was slightly larger than pre-procedure, enhancement scan showed the thin homogeneous ring enhanced around the non-enhanced lesions; 4 months later, the size of lesions were relative stable and the periphery enhancement was weaken; 7 months later, the size of lesions were reduced and showed no enhancement. For recurrence lesions, the high signal ring was incomplete on TIWI, the incomplete area showed nodular enhancement on the arterial phase, and most of nodule showed slightly lower signal on the delay phase demonstrated a feature of quick wash-in and wash-out; 7 months after RFA, recurrence rate was 6.12% in patients with complete high signal ring and 43.75% in patients with incomplete high signal ring, the difference was statistically significant (P<0.05). The total survival rate and accumulated survival rate of the patients with complete high signal ring on T1WI were higher than the patients with incomplete ring, the difference was statistically signiifcant (P<0.05).ConclusionThere are characteristic ifndings of MRI examination of liver cancer after percutaneous RFA, observation of the integrity of high signal ring on T1WI image and ifnding of dynamic enhancement scan can early evaluate efifcacy of RFA guide the selection of treatment plan.

AIM: To specify optimization of the concentration, purification and drying for prepar ati on of Yuxianling Grannles and decrease the dosage on condition that is retentive of potency in order to provide a basis for mass production. METHODS: The contents of ferulic acid and the amount of extract wer e used as marker, the optimium of concentration, purification and drying for pr eparation were selected by orthogonal design and contrast test during aqueous ex tract alcohol precipitation process. RESULTS: Condition was optimum of herb-to-extract ratio in the range of 0.5 ∶1, pr ecipitation with ethanol containing 40% water and drying temperature not to exce ed 60℃ . CONCLUSION: The highest yield of active priciples is obtained by c ondition above.

OBJECTIVETo investigate the anti-proliferative effect of Protobioside on the HepG2 cells and its mechanism.

METHODThe inhibitory effects of Protobioside on 3 kinds of human cancer cell line was measured by MTT assay. Apoptotic morphological changes was observed by Hoechst33528 staining technique. Cell cycle were analyzed using flow cytometry. The expression of cell cycle related protein and apoptosis related protein were determined using Western blotting.

RESULTProtobioside shows strong cytotoxic power on HepG2 cells and IC50 were 20 micromol x L(-1). The nucleus became pyknosis at 36 hours. The cells in G2/M phase increase in a dose-dependent and time-dependent manner. And the protein level of CyclinB1 was down-regulated, that of Bax was up-regulated and that of Bcl-2 was down-regulated.

CONCLUSIONThe growth inhibition of Protobioside on HepG2 cells is highly related to cell cycle arrest at G2/M phase which was well correlated with the down-regulation of expression of Cyclin B1, and the induction of apoptosis via up-regulating of Bax and down-regulating Bcl-2 expression.

Apoptosis ; drug effects ; Blotting, Western ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Hep G2 Cells ; Humans

Objective To characterize clinical feature, frequency of isolation and antimicrobial susceptibility of pathogens isolated from patients with nosocomial bloodstream infections in Huashan Hospital, Fudan University from 1995 to 2004. Methods The clinical data of all patients who were diagnosed with nosocomial bloodstream infections based on national diagnostic criteria of nosocomial bloodstream infections were retrospectively analyzed. The pathogens were routinely isolated and identified. Susceptibilities against antimicrobial agents were determined by Kirby-Bauer methods and analyzed by WHONET 5.0 software. Results During the 10-year study period, a total of 395 patients were diagnosed with nosocomid bloodstream infection with 435 strains isolated from blood specimen.Gram positive bacteria, Gram negative bacilli and fungi, accounted for 47.4%, 45.1 % and 7.6%,respectively. Coagulase-negative Staphylococci (21.4%), S. aures (17.9%), E.coli (13.6%), K. pneumoniae (10.8%), Candidaspp (7.4%), Enterococci (6.0%), Pseudomonasspp (6.0%) and Acinetobacter spp (3.7%) were frequently identified isolates. S. aures and coagulase-negative Staphylococci resistant to methicillin were 62.8% and 87.1%, respectively. The susceptibilities of cefotaxime and ceftazidime against E. coli and K. preumonine were 46%-78% and 27.7%-40.4%, respectively. Conclusions The Gram positive cocci are slightly more prevalent than Gram negative bacilli in nosocomial bloodstream infections and resistance to the first line antibiotics is common among all pathogens isolated. Candida spp is the fifth leading cause of nosocomial bloodstream infections.

AIM: Anti-atherosclerosis effects of Momordica charantia L was further studied in a New Zealand rabbit atherosclerotic model at the basis of anti-inflammation and antioxidant effects. METHODS: Animals were divided into 3 groups: normal group (normal rabbit diet), atherosclerosis group(diet containing 2% cholesterol), and Momordica charantia L group(diet containing 2% cholesterol and 1 5% sarcocarp of Momordica charantia L ). Ninety days later, all animals were sacrificed. The effect of Momordica charantia L on atherosclerosis was evaluated by measuring serum lipid and total cholesterol content of artery wall, observing fatty liver degree, aorta arteriosclerotic area, and the thickness of intima. RESULTS: The level of total serum cholesterol and LDL-C in Momordica charantia L treatment group were obviously lower than those in atherosclerosis group, so were the total cholesterol content of artery wall, fatty liver degree, atherosclerotic area, intima thickness and I/M ratio, but no significantly difference was found between the two groups in TG level. The level of HDL-C in Momordica charantia L treatment group was evidently lower than that in normal control group. CONCLUSION: Momordica charantia L has an anti-atherosclerosis action in rabbits.

OBJECTIVETo study on pharmacologic actions on quail hyperlipidemia and atherosclerosis model.

METHODTo duplicate quail hyperlipidemia model by ectogenesis cholesterol and high fat forage, induce to atherosclerosis model, observe influence of sugarcane alkane alcohol to model animals' blood fat level, formation of atherosclerosis plaque, pathological changes of coronary vessels and vascular intimal.

RESULTTC, TG, LDL-C level in blood serum of quail hyperlipidemia markedly decreased after administered sugarcane alkane alcohol by dose of 30, 15, 7.5 mg x kg(-1), proliferation of aorta and brachiocephalic artery tunica intima foam cells was suppressed.

CONCLUSIONSugarcane alkane alcohol has satisfactory pharmacologic actions on hyperlipidemia and atherosclerosis animal model by regulating blood fat.

Alkanes ; chemistry ; therapeutic use ; Animals ; Atherosclerosis ; drug therapy ; Cholesterol ; blood ; Disease Models, Animal ; Ethanol ; chemistry ; therapeutic use ; Hyperlipidemias ; drug therapy ; Male ; Plant Extracts ; chemistry ; therapeutic use ; Quail ; Random Allocation ; Saccharum ; chemistry

Background and purpose:Urothelial carcinoma of the bladder (UCB) is the most common cancer in urinary system. Yes associated protein (YAP) gene is closely associated with urothelial carcinoma of the bladder. The study was aimed to explore the effect of siRNA targeting the YAP gene on cell proliferation and migration of T24 cells. Methods:Small interfering RNA (siRNA) was transfected together with LipofectamineTM2000 in T24 human bladder cancer cells to block the YAP signal pathway. The effect of siRNA on cell proliferation and invasiveness was assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay. Quantitative real time-Polymerase chain reaction (qRT-PCR) and Western blot analysis were used to conifrm the successful suppression of YAP gene and protein by siRNA. Results:Expression of YAP gene and protein was successfully suppressed after transfected with siRNA which verified by qRT-PCR and Western blot(RNA:F=93.91, P<0.000 1; Protein: F=4.62, P<0.05). As CCK-8 test showed, the proliferation of T24 bladder cancer cells was successfully restrained by inhibition of YAP gene compared with blank control and negative control(12 h: F=6.00, P=0.037;24 h: F=41.72, P=0.000 3;36 h:F=462.8, P<0.000 1;48 h:F=236.6, P<0.000 1;72 h:F=140.5, P<0.000 1). Transwell and wound healing test were performed after YAP gene was interfered by siRNA. The result demonstrated that migration of T24 bladder cancer cells was signiifcantly inhibited (Transwell: F=43.55, P<0.05;Wound healing: F=43.55, P<0.05). Conclusion:This study suggested that YAP gene was an important enhancer for the proliferation and migration of bladder cancer cells.

OBJECTIVETo study chemical constituents contained in Desmodium caudatum.

METHODThe chemical compounds were separated by using such chromatographic methods as macroporous resin, Sephadex LH-20, ODS and normal phase silicagel column, and their structures were identified by spectroscopic data analysis.

RESULTFifteen compounds were separated and identified as stigmasterol (1), beta-sitosterol (2), citrusinol (3), hibiscone A (4), yukovanol (5), kenusanone I (6), neophellamuretin (7), desmodol (8), erythrotriol (9), hibiscone D (10), kaempferol (11), 8-prenylquercetin (12), leachianone G (13), 5,7,4'-trihydroxy-dihydroflavonol (14), and 4H-1-benzopyran-4-one, 2-(3,4-dihydroxyphenyl) -2, 3-dihydro-3,5,7-trihydroxy-8-( 3-methyl-2-butenyl) -, (2R-trans)-(9CI) (15).

CONCLUSIONAll of the compounds were separated from D. caudatum for the first time except compound 8.

Drugs, Chinese Herbal ; chemistry ; Fabaceae ; chemistry ; Organic Chemicals ; analysis ; isolation & purification ; Spectrum Analysis

With the discovery of exosomes,new pathways of intercellular information and material exchange mediated by exosomes are attracting more and more attention from researchers. The process of exo-some production overlaps with many viral assembly and outflow pathways,suggesting that exosomes may be related to viral infections. In vitro experiments also show that exosomes play a very important role in viral in-fections. On one hand,exosomes can transfer viral nucleic acids and proteins,and may change microenviron-ment to promote the spread of infection. On the other hand,exosomes can induce immune responses by activa-ting antiviral pathways or transferring antiviral molecules. Do they promote or suppress the spread of infec-tion? What are the factors that affect their functions? In this paper,we review the role of exosomes in viral in-fection in order to provide a reference for better understanding the process of viral infection and immune re-sponses,and to provide a new train of thoughts for the prevention,diagnosis and treatment of viral diseases.