Objective To establish an accurate method for determining the content of three components in Fufang Buwu Syrup. Methods TLC scanner was selected to detect three components with silica gel G thin layer plate. The sample was separated by using cyclohexane-ethyl acetate-methylenechloride-formic acid (3∶1∶1∶0.2),λS=300 nm. Results The linearity between peak area and ferulic acid was achieved in the range of 0.36-0.84μg, psoralen was achieved in the range of 0.12-0.28 μg, emodin was achieved in the range of 0.01-0.05 μg. The average recovery was 100.7%, 100.8%, 101.0%, and RSD was 1.26%, 1.44%, 1.86%, respectively. Conclusion The method is simple and accurate, which can be used for quality control of Fufang Buwu Syrup.

Aim To investigate the effect of irbesartan and spironolactone on expressions of connective tissue growth factor(CTGF),P-p38MAPK proteins during vascular remodeling in renovascular hypertensive rats(RHR).Methods Renovascular hypertension was induced by two kidney one clip(2K1C)operation.8 weeks later,RHRs were given irbesartan or(and)spironolactone for 8 weeks.After 8 weekstreatment,the morphometric measurements were performed in the mesenteric arterioles.Concertration of carboxyterminal propeptide of typeⅠprocollagen(PⅠCP)and N-terminal propeptide of type Ⅲ procollagen(PⅢNP)in blood serum was measured by enzyme linked immunosorbent assay method,and the media collagen area was assessed by collagne-specific Picro-sirius red staining with computerized video processing.Expressions of collagen type I,CTGF,P-p38MAPK proteins were detected using immunohistochemistry respectively.Results The arterial systolic pressure was attenuated significantly after the treatment of irbesartan,and this effect could not be enhanced by spironolactone.The media thickness over lumen ratio,media cross-sectional area over luminal area ratio of mesenteric arterioles,concertration of PⅠCP and PⅢNP,media collagen area,and expression of collagen typeⅠwere significantly increased in RHRs but ameliorated by irbesartan and spironolactone.Meanwhile,the expressions of CTGF,P-p38MAPK proteins were up-regulated in RHRs but blunted significantly after the treatment of irbesartan and spironolactone.The combined treatment had the synergic effects.Conclusions There is a synergistic effect of attenuating extracellular matrix(ECM)producing and amelioration of vascular remodeling(VR)by combined use of irbesartan and spironolactone.It maybe related to the expressions of CTGF and P-p38MAPK proteins down regulated by these two drugs.

BACKGROUND: The deficiency of perfect animal femoral head necrosis model limited its further investigation. OBJECTIVE: To verify the feasibility of establishing rabbit femoral head necrosis models using liquid nitrogen rsfdgeration method, and to provide a foundation for subsequent research. METHODS: A total of 20 adult, New Zealand, white rabbits were selected in the study. The round ligament of femur was not cut off and femoral head was not dislocated, and the exposed femoral head were quick frozen using cotton bud carrying liquid nitrogen for successive 25 times, with 10 s per time. The specimens were examined by gross anatomy, X-ray film, MRI and histological observation at day 3, 7 and weeks 2, 4, 6, and 8 after operation. RESULTS AND CONCLUSION: The histolOgical section showed that chondrocyte, osteccyts, and myelold tissues presented necrosis in freezing and periphery at 3days after model preparation, and the repair process appeared at 2weeks after operation. The articular surface of femoral heads appeared collapse at 4 weeks after operation, and these changes became obvious at 6 weeks. The femoral head presented ostecarthdtis-like disorder, with seriously collapsed articular surface at8 weeks, and the contour of femoral head changed in 2 animals. The results demonstrated that without hip dislocation, rabbit femoral head necrosis models can be established successfully using liquid nitrogen refrigeration method. This method is simple, feesible, with high succeed rate, which can be used in subsequent research.

BACKGROUND: The freeze-drying blood vessel is an excellent material for vasotransplantation. However, the reports concerning freeze-drying are few, and the mechanical property changes of freeze-dried blood vessel remains poorly understood. OBJECTIVE: Using micro-CT to explore the morphological changes of blood vessel during procedure of freeze-drying. DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Refrigeration and Cryogenics, University of Shanghai for Science and Technology. MATERIALS: Fresh pig aorta was obtained from Shanghai Slaughter House. The lyophilizer was produced by SP Industries (USA). TRAPEZIUM LITE texture analyzer was purchased from Shimadzu (Hong Kong) Co., Ltd.METHODS: The differences of mechanical property between fresh aorta and freeze-dried aorta were compared.MAIN OUTCOME MEASURES: ① Temperature curve of freeze-drying procedure. ②The reeze-drying procedure was scanned by micro-CT. ③ Changes of mechanical property were detected by TRAPEZIUM LITE texture analyzer and puncture experiments.RESULTS: During freeze-drying procedure, there were presented layer phenomenon in pig aorta. Compared to the fresh aorta, the freeze-dried aorta after recover water would reduce 40% axial tensile force, increase 45% circumferential tensile force, with 75% puncture stress.CONCLUSION: The freeze-dried blood vessel maintains mechanical property as fresh vessel, thus, freeze-drying can be used for vascular preservation.

Aim To observe the effects of benazepril and irbesartan on myocardial collagen in chronic heart failure ( CHF ) rats and the possible mechanisms. Methods CHF model in rats was made by partially banding abdominal aorta to induce pressure overload cardiac hypertrophy. The rats were given benazepril or/and irbesartan for 8 weeks. Systolic blood pressure ( SBP) was monitored by rat tail artery. The ultrastruc-ture of myocardium was detected by transmission elec-tron microscope. The content of myocardial hydroxyproline and amount of cross-linked collagen were measured by hydrolysis method. The expression of type Ⅰ and Ⅲ collagen protein in myocardium was investigated by immunohistochemistry. The expression of p-38 MAPK and p-p38 MAPK proteins was detected by Western blot. Results Compared with the model of CHF, there was no significant difference in SBP af-ter treatment. The content of hydroxyproline in myocar-dium, degree of cross-linked collagen, as well as ex-pression of type I collagen, type Ⅲ collagen and p-p38 MAPK proteins were significantly decreased after treatment with benazepril or irbesartan ( P <0. 05 ) , and the combined treatment of them had better effects. Conclusion There is a synergistic effect of attenua-ting the quantity and quality of myocardial collagen in CHF rats by combined use of benazepril and irbesar-tan, which may be related to the down regulation of p-p38MAPK protein expression.

OBJECTIVEIt has been known that the intrahepatic renin-angiotensin-aldosterone system (RAAS) plays a key role in the fibrogenesis in livers. Aldosterone (Aldo), the principal effector molecule of the RAAS, exerts local effects on cell growth and fibrogenesis. However, the signal transduction mechanisms underlying the effects of Aldo on hepatic fibrogenesis remain to be fully elucidated. The present study aims to investigate the signal transduction mechanism underlying the effects of Aldo on the signal passageway of active protein-1 (AP-1).

METHODSIn vitro, HSCs-T6 cell line was treated with Aldo for 10 min, 30 min, 60 min, 120 min and 180 min, and protein expression of Phospho-P42/44 was detected by Western blot. In addition, HSCs-T6 cell line was preincubated for 60 min or not with U0126 (an inhibitor of the MAPK/ERK kinase), and also with antioxidant-N-acetylcysteine (NAC) prior to exposure to Aldo for the indicated times. Protein expression of Phospho-P42/44 was measured by Western blot. DNA biding activity of AP-1 was analyzed by electrophoretic gel mobility shift assay (EMSA). By means of RT-PCR, expression of alpha1(1) procollagen mRNA was detected.

RESULTSAldo stimulated HSC via extracellular signal-regulated kinase (ERK1/2) pathway. Time course experiments showed that Aldo induced Phospho-P42/44 expression, which was abrogated by U0126, reaching a maximum at 10 minutes, and then declined progressively. NAC inhibited the Phospho-P42/44 expression. EMSA showed that stimulation of HSC by Aldo markedly increased AP-1 DNA binding activity. U0126 markedly reduced AP-1 DNA binding activity induced by Aldo; NAC partly decreased AP-1 activity induced by Aldo. Aldo up-regulated expression of alpha1(1) procollagen mRNA, which was attenuated by U0126 and NAC.

CONCLUSIONStimulation of HSC by Aldo results in activation of AP-1 via ERK1/2 pathway, leading to up-regulation of AP-1 target gene alpha1(1) procollagen mRNA expression.

Aldosterone ; pharmacology ; Cell Line ; Collagen Type I ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 3 ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Transcription Factor AP-1 ; metabolism

Adult ; China ; epidemiology ; GB virus C ; classification ; Hepatitis, Viral, Human ; epidemiology ; virology ; Homosexuality, Male ; Humans ; Male

Objective To evaluate the setup errors of image guided radiation therapy (IGRT) for head-and-neck cancer using kilovoltage cone beam CT( kV CBCT).Methods 256 patients with head-and-neck cancer were treated with intensity modulated radiation therapy (IMRT) from March 2009 to October 2011.All patients were immobilized with head-and-neck mask and localized with final isocenter marking method using the Philips PQS CT or Philips Brilliance CT Big Bore scanners,which were equipped with LAP movable laser systems.The CT images were transferred to a Varian Eclipse V8.6 workstation for contouring and planning.A kV cone-beam CT scans was acquired,and registered before the treatment for every patient on a Varian iX linear accelerator via OBI system.The setup errors in the right-left ( RL),superior-inferior (SI),and anterior-posterior (AP) directions were recorded.Results The setup errors for the 473 datasets followed a Gaussian distribution.The systematic errors ± random errors in the RL,SI and AP were(-0.6 ± 1.3 ),(0.5 ± 1.6) and (0.9 ± 1.7 ) mm,respectively.The planning target volume (PTV) margins were calculated respectively as 2.4,2.4 and 3.4 mm according to the formula of M =2.5∑ +0.7δ The margins of 288 sets of data using the Big Bore CT scanner were calculated as 2.0,2.1 and 1.7 mm,respectively.Conclusions The setup errors using final isocenter marking method are smaller than those using reference point marking method.The result derived from this retrospective study could be used to set the margin between CTV and PTV.

Objective: To explore the application and affecting factors of clopidogrel loading therapy in patients with ST-segment elevation myocardial infarction (STEMI) undergoing percutaneous coronary intervention (PCI) in china in 2006 and 2011. Methods: Based on “China patient-centered evaluative assessment of cardiac events retrospective study of acute myocardial infarction”, we collected clinical information of STEMI patients with PCI in 2006, 2011 and analyzed clopidogrel loading therapy status in different years and different hospitals. According to clopidogrel doses, the patients were divided into 2 groups: Clopidogrel 300mg group, the patients received a single dose of clopidogrel ≥300mg while <450mg and Clopidogrel 600mg group, the patients received a single dose of clopidogrel ≥450mg while ≤600mg. The relevant factors affecting clopidogrel loading therapy status were identiifed by binary Logistic regression analysis. Results: A total of 2481 eligible patients were enrolled and their mean age was (60.9 ± 12.0) years including 21.4%female. From 2006 to 2011, the overall application of clopidogrel loading was (51.0% vs 47.4%), the ratio for patients in Clopidogrel 300mg group was (43.1% vs 39.2%), in Clopidogrel 600mg group was (7.8% vs 8.2%), allP>0.05. In 2006, the hospital median rate of clopidogerl application was 44.4% (IQR 21.8% to 69.0%) and in 2011, it was 48.1% (IQR 25.0% to 70.8%),P=0.940. Binary logistic regression analysis showed that the patients were admitted within 12 hours of onset, with primary PCI and treated in central region had the higher rates of clopidogrel loading therapy. Conclusion: Clopidogrel loading therapy was seriously inadequate in STEMI patients with PCI, variation was across hospitals and the status was similar between 2006 and 2011. Clopidogrel loading therapy should be improved.

Objective: To explore the thermal damage to epithelial cell adhesion molecule(EpCAM)-positive tumor cells by novel aptamer-guided magnetic nanoparticles(AptNPs). Methods: EpCAM aptamer SYL3C was connected to NPs via biotin-streptavidin reaction. The diameter of AptNPs were characterized by Dynamic Light Scattering(DLS). The binding feature of the aptamer to EpCAM-positive tumor cells was evaluated by Prussian blue dyeing. Thermal damage under alternative magnetic field was measured bylactate dehydrogenase (LDH). The apoptosis of EpCAM-positive tumor cells was detected by acridine orange/ethidium bromide (AO/EB) double staining. Results: The average size of AptNPs was 282 nm. Flow cytometry and Prussian blue dyeing showed that AptNPs exhibited strong binding to the EpCAM-positive tumor cells but not to the EpCAM-negative tumor cells. Moreover, when incubated with 1.5×10⁸ AptNPs under alternative electromagnetic fieldfor 5 hours, the viability of EpCAM-positive HCT116 cells and A549 cells was 28.9% and 54.4%, respectively, significantly lower than 76.7% of EpCAM-negative HepG2 cells (P<0.05). Conclusions AptNPs can improve the thermal damage to EpCAM-positive tumor cells, and may have potential utility in the development of tumor targeted therapy.