Objective To explore the effect of different antifreeze or antioxidant at different cryopreservation conditions on the activity and the morphology of adipose cells.Methods After purification,fat granules were divided into 5 groups; the same volume of antifreeze or antioxidants were added in each group,stored at-20℃ (refrigerator),-80℃ (ultra-low-temperature refrigerator) and-196 ℃ (liquid nitrogen) ; the amount of glucose transfer was determined and the morphological changes were observed for each group after two weeks,two months,and three months.Results Fat activity of the group adding both dimethyl sulfoxide (DMSO) and trehalose was significantly higher than that in other groups at the same temperature and the same time point.The decreased amount of glucose transfer of 80℃ group and 196℃ group was significantly lower than that of 20℃ group,and that in groups of each 196℃ were greater than that in groups of each 80℃.Conclusions Antifreeze can obviously keep activity of fat cells.Preservative effect of DMSO is better than trehalose,and that of both combination is much better.Liquid nitrogen storage of adipose tissue for three months and fat granules stored at 80℃ for two months are still suitable for clinical use.

Objective To investigate the expressions of high mobility group box-1(HMGB1) and high sensitivity C-re?active protein (hs-CRP) in patients with acute coronary syndrome (ACS) and the effects of atorvastatin on the two inflamma?tory cytokines. Methods A total of 90 patients with ACS and 90 cases of normal control subjects were selected in this study. The serum concentrations of HMGB1 and hs-CRP were measured before treatment in patients of ACS. Patients were randomly divided into two groups:control group (n=45) and atorvastatin group (n=45). Atorvastatin was given 20 mg/24 h and 40 mg/24 h. Blood samples were obtained from the patients for detection of HMGB1 and hs-CRP one week after treatment with atorvastatin. Results There were significantly higher serum levels of HMGB1 and hs-CRP in patients with ACS than those of control subjects (P<0.01). The level of HMGB1 was positively correlated with the level of hs-CRP in patients of ACS (r=0.389, P<0.01). Before treatment, there were no significant diffferences in level of HMGB1 and hs-CRP in patients with ACS between the two groups. After treatment with atorvastatin, the levels of HMGB1 and hs-CRP were decreased in the two groups of ACS, and those were significantly lower in the intensive group than the standard group (P<0.05). Conclu?sion HMGB1 could stimulate the secretion of hs-CRP and other inflammatory cytokines, playing an important role in the process of occurrence and development of atherosclerosis. High loading dose of atorvastatin may reduce the expression of HMGB1 and decrease the inflammation, and stabilize the plaques in patients with acute coronary syndrome.

Objective To investigate the effects of high loading dose of atorvastatin on lipoprotein-associated phospho?lipase A2 (Lp-PLA2) and inflammatory cytokines in patients with acute myocardial infarction (AMI), who underwent emergen?cy percutaneous coronary intervention (PCI). Methods A total of 65 cases with AMI who underwent emergency PCI be?tween October 2011 and August 2013 were randomly divided into two groups:control group (n=32, atorvastatin 20 mg/24 h) and high dose atorvastatin group (n=33, atorvastatin 40 mg/24 h). Two groups of patients were given the same basic treat?ment. Blood samples were obtained before treatment and 72 h after PCI in two groups. Levels of Lp-PLA2, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), alanine transaminase (ALT) and aspartate transaminase (AST) were detected in two groups. The adverse drug reactions were observed. Results There were no significant differences in Lp-PLA2, IL-6, TNF-α, ALT and AST levels between two groups (P>0.05). After PCI, the levels of Lp-PLA2, IL-6 and TNF-αwere significantly increased compared with those of baseline in two groups, and they were more notable in control group than those of high dose atorvastatin group (P<0.05). The levels of ALT and AST were also significantly higher after operation compared with those of basic levels (P<0.05), while there were no significant differences in ALT and AST after PCI between two groups (P>0.05). Conclusion The high loading dose of atorvastatin in AMI patients underwent emergency PCI can de?crease the inflammation and stabilize the plaques in acute stage, and which is safe.

Objective To investigate the effects of perioperative high loading dose of Atorvastatin treatment on lipoprotein associated phospholipase A2 (Lp-PLA2) and heart function in patients with acute ST-segment elevation myocardial infarction and type 2 diabetes who underwent emergency percutaneous coronary intervention (PCI).Methods Totally 83 cases with acute ST segment elevation myocardial infarction and type 2 diabetes who underwent emergency PCI from September 2012 and August 2014 were randomly divided into two groups.In control group (n=42)patients took Atorvastatin 20 mg daily before and after emergency PCI,and in intensive group (n=41) patients took atorvastatin 40 mg daily before and after emergency PCI.Each group was given the same basic treatment according to the guideline.Blood samples were obtained from all the patients before PCI and at 3,7 days after PCI,and levels of Lp-PLA2 and brain natriuretic peptide (BNP)were detected.And the left ventricular end diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) were measured at 1 day and 1 month after PCI.Results The levels of Lp-PLA2 and BNP at 3 days after PCI were obviously increased in the two groups versus baseline [(297.8± 53.4) mg/L vs.(194.7±39.1) mg/L,(270.3±47.0) mag/L vs.(205.6±27.5) mg/L,both P＜0.05],and decreased in intensive versus control group [(270.3±47.0) mg/L vs.(297.8±53.4)mg/L and (353.8±76.3) mg/L vs.(375.4±57.0) mg/L,P＜0.05].And levels of Lp-PLA2 and BNP at 7 days after PCI were improved more in intensive than in control group [(227.2±33.3)mg/L vs.(249.3±42.3) mg/L,(206.0±48.2)mg/L vs.(267.6±50.8) mg/L,P＜0.05].There were no significant differences in LVEDD and LVEF between the two groups 1 day after PCI.Meanwhile,the LVEDD was decreased and the LVEF was increased in the two groups 1 month after PCI as compared with 1 day after PCI (both P＜0.05).Conclusions Perioperative high loading dose of Atorvastatin treatment may stabilize the plaques and improve heart function in acute stage in patients with acute ST-segment elevated myocardial infarction and type 2 diabetes after emergency PCI.

Objective To investigate the effects of ibutilide and amiodarone on the ventricular transmural heteroge-neity of repolarization and ventricular arrhythmia for the treatment of atrial fibrillation. Methods Eighty-seven patients with paroxymal atrial fibrillation at 48 h~7 d were enrolled and randomized to two groups, ibutilide and amiodarone treat-ment groups. The successful rate of cardioversion to sinus rhythm was compared between two groups. The electrocardiograph-ic QT interval and Tpeak-end/QT ratio were also analyzed before and after treatment in two groups. Results The successful rate of cardioversion was significantly higher in ibutilide group than that of amiodarone group (61.7%vs 40.7%, P＜0.05). The QT intervals and Tpeak-end/QT ratio were both significantly increased in ibutilide group (P＜0.05), which were re-turned to the levels before treatment in 2 hours and 1 hour, respectively (P＜0.05). The QT intervals were significantly in-creased in the amiodarone group (P＜0.01), which were continued until 4 h after treatment. There were no significant differ-ences in the Tpeak-end/QT ratios before and after treatment (P>0.05). Conclusion The successful rate of cardioversion to sinus rhythm for atrial fibrillation by ibutilide was significantly higher compared with that of amiodarone. Ibutilide slightly in-creased the transmural heterogeneity of repolarization within the first hour, which may increase the risk of ventricular arrhyth-mia.

Directive 2004/24/EC of the European Parliament and the Council entered into force on April 30th, 2004. After 7 years, there is no Chinese medicine to be registered successfully in European market as traditional herbal medicinal products. The thesis gives some ideas to tackle this problem. Procedure for the Preparation of Community Monographs for Traditional Herbal Medicinal Products (EMEA/HMPC/182320/2005) published by European Medicines Agency is an important guidance for traditional herbal products to enter European Community monographs. The thesis introduces and details the procedure as well as gives feasible suggestions about the procedure. It suggests that Chinese medicines enter European Community monographs first, and then apply the registration according to the directive 2004/24/EC. This is an easier access to European market.

ObjectiveTo investigate the mechanism of Yiqi Huoxue Tongluo prescription （YHTP） in the treatment of diabetic neuropathic pain （DNP）. MethodNinety SPF-grade SD male rats were randomized into blank， model， low- （2.25 g·kg^-1）， medium- （4.5 g·kg^-1）， and high-dose （9 g·kg^-1） YHTP， and mecobalamin （0.175 mg·kg^-1） groups. Except those in the blank group， the rats in the remaining 5 groups were fed with a high-fat and high-glucose diet and subjected to intraperitoneal injection of low-dose （35 mg·kg^-1） streptozotocin （STZ） to establish the model of DNP. The sciatic nerve conduction velocity in DNP rats was measured by the neurophysiological method， and the levels of interleukin-6 （IL-6）， IL-1β， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to measure the mRNA levels of glial fibrillary acidic protein （GFAP） and extracellular signal-regulated kinase （ERK） in the spinal cord. Western blot was employed to measure the protein levels of GFAP and phosphorylated ERK （p-ERK）， and immunofluorescence staining to measure the fluorescence intensity of GFAP and p-ERK in the spinal cord. In the cell experiments， 100 mmol·L^-1 high glucose was used to induce the activation of astrocytes （CTX-TNA2） for the modeling of nerve cell injury. The cells were randomized into the normal， model， drug-containing serum （10% YQHT）， inhibitor ［10 mol·L^-1 corynoxeine （COR）］， drug-containing serum + inhibitor （10% YHTP + 10 mol·L^-1 COR） groups. The levels of pro-inflammatory factors （TNF-α and IL-1β） and the anti-inflammatory factor IL-10 in CTX-TNA2 cells were determined by ELISA， and the protein levels of GFAP and p-ERK in CTX-TNA2 cells by Western blot. ResultThe animal experiments showed that compared with the blank group， the model group presented reduced mechanical withdrawal threshold （MWT）， thermal work limit （TWL）， and nerve conduction velocity， elevated levels of fasting blood glucose， IL-1β， TNF-α， and IL-6， and up-regulated protein levels of GFAP and p-ERK， and mRNA levels of ERK1， ERK2， GFAP （P<0.01）. Compared with model group， YHTP increased the MWT， TWL， and sciatic nerve conduction velocity （P<0.01）， lowered the levels of IL-1β， TNF-α， and IL-6 （P<0.01）， and down-regulated the protein levels of GFAP and p-ERK， and mRNA levels of ERK1， ERK2， GFAP in the spinal cord （P<0.05， P<0.01）. The cell experiments showed that compared with the blank group， the model group had decreased survival rate， elevated levels of pro-inflammatory factors， and up-regulated protein levels of ERK and GFAP （P<0.01）. Compared with the model group， the YHTP-containing serum lowered the levels of IL-1β and TNF-α （P<0.05， P<0.01）， elevated the level of IL-10 （P<0.01）， and down-regulated the protein levels of ERK and GFAP （P<0.01）. ConclusionYHTP may inhibit the activation of astrocytes by inhibiting the ERK signaling pathway to reduce inflammation and thus relieve DNP.