The aim of this study is to improve liposome encapsulation efficiency of water soluble drug ATP-2Na with hydrophobic ion pairing method, and evaluate its effect on tissues energy state in myocardial ischemia mice. Ion pair complex of ATP-2Na with HTAB was prepared first; then the liposomes were prepared by ethanol injection method. The size and zeta potential of ATP-2Na liposome were investigated. Its effect on tissues energy state in myocardium ischemia mice was evaluated by detecting ATP-2Na concentration in tissues and blood after injection in comparison to ATP-2Na solution. The diameters and zeta potential of ATP-2Na liposomes were (144.0 +/- 2.7) nm and (+16.2 +/- 1.6) mV, respectively. The encapsulation efficiency was (85.02 +/- 2.31) %. The in vitro drug release pattern from liposomes matches with Weibull equation. Compared with ATP-2Na solution, ATP-2Na liposome increased the ATP concentration of blood in myocardial ischemic mice very significantly; compared with blank, ATP-2Na liposome increased ATP content of myocardium and liver in myocaridal ischemic mice significantly, but ATP-2Na solution didn't show this effect. ATP-2Na liposome might have an advantage in improving tissue energy state on myocaridal ischemic animals.
Adenosine Triphosphate ; administration & dosage ; blood ; metabolism ; Animals ; Cetrimonium Compounds ; chemistry ; Drug Carriers ; Liposomes ; chemistry ; Liver ; metabolism ; Male ; Mice ; Myocardial Ischemia ; blood ; metabolism ; Myocardium ; metabolism ; Particle Size ; Random Allocation ; Surface-Active Agents ; chemistry

Background Infective keratopathy is a key cause of corneal blindness in China,and fungal keratitis is proved to have a higher incidence and bigger threats in infective keratitis.Researches showed that topical immunology plays an important effect during the development of fungal keratitis,but its mechanism is still studying.Objective This experiment was to explore the critical immunocyte during the process of fungal keratitis.Methods Forty-eight SPF 12-week-old male C57BL/6J mice were included and randomized into the control group and model group.The fungal keratitis model closely mimicking human cornea infections was established in the mouse using scratch followed by incubation of fusarium solani on the cornea,and the mice in the control group scratched on the cornea only.Cornea was examined under the slit lamp at 0,6,9,12,24,72 and 120 hours after operation.The severity of keratomycosis was clinically scored based on the literature criteria.The inflammatory cells were identified using immnofluorescence label,and the number of the inflammatory cells was calculated and compared among different groups and time points.This study complied with the Statement of ARVO in the use of experimental animal.Both Experimental Animal Ethic Commission in Zhengzhou University and Life Science Management Commission approved this study proposal.Results After inoculation of fusarium solani,typical fungul keratitis signs were seen on the cornea.Severe corneal opacifieation occurred within 24 hours and peaked at 72 hours.However,only mild edema of cornea was exhibited and gradually recovered normal in the control group within 24 hours.The clinical score of inflammation was higher in the model group in various time points than that in the control group,and it was seen that 24-72 hours after operation,the score attached peak in the model group with a significant difference in comparison with the control group(P＜0.01).In 9,12,24,72 and 120 hours after operation,the number of neutrophil cells was significantly increased in the model group compared with control group (P＜0.05),and that in 12,24,72 hours after operation was significantly higher than the 6 hours(P=0.004,0.000,0.001).However,no significant differences were seen in the number of neutrophil cells between 9 or 120 hours and 6 hours after operation(P=0.772,0.323).The number of T lymphocytes in cornea was significantly increased in 72 and 120 hours in comparison with 6 hours in the model group(P=0.000,0.000),and from 72 to 120 hours after operation,the number of T lymphocytes was significantly higher than that of the contral group (P＜0.01).The neutrophil cell number was positive correlated with the inflammatory score in the early phase (r =0.593,P =0.000).T limphocyte emerged in late phase but no significant correlation with the clinical score (r＝0.315,P=0.062).Conclusions Neutrophil cells play a critical role in the development of fungal keratitis in early stage.

OBJECTIVETo explore the mechanisms of a novel potassium channel gene named KCTD9 (potassium channel tetramerization domain containing 9) in model of fulminant viral hepatitis induced by murine hepatitis virus 3 (MHV-3).

METHODS78 BALB/cJ mice(6 male) were randomly and equally assigned to two groups, model group of fulminant viral hepatitis induced by MHV3 and its control. 75 C3H/HeJ female mice were done into two groups, 39 for model group of chronic hepatitis induced by MHV3, 36 for control. Various samples including spleen, liver and lymphocytes from mice of two model groups and the controls were examined for KCTD9 expression by real time quantitative PCR and Immunohistochemistry. Independent-samples T test or one-way ANOVA were carried out in different groups.

RESULTSIncreased expressions of KCTD9 mRNA was observed in livers of both model mice of fulminant viral hepatitis and chronic hepatitis. Compared with the control mice, the expressions of KCTD9 mRNA were up-regulated by 577.1-, 8.8-, 59.4- and 10.8-fold in hepatic NK cells, CD4+ T cells, CD8+ T cells and splenic NK cells respectively in model mice of fulminant viral hepatitis 48 hr post MHV-3 infection, whereas down-regulation by 43% and 69% in splenic CD4 + T cells and CD8+ T cells were found respectively. In contrast, in model mice of chronic viral hepatitis the expressions of KCTD9 mRNA were down-regulated by 71% and 51% in hepatic CD4+ T cells and NK cells, respectively. The expression of KCTD9 protein was mainly evidenced in infiltrative mononuclear cells of liver as shown by immunohistochemistry. Basal expression was also investigated and showed constitutive expression of KCTD9 in brain, thymus and other organs in BALB/cJ mice.

CONCLUSIONA novel potassium channel gene KCTD9 was highly expressed in hepatic NK cells and T cells of fulminant hepatitis mice induced by MHV-3.

Animals ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; Female ; Hepatitis, Viral, Animal ; immunology ; metabolism ; virology ; Killer Cells, Natural ; immunology ; metabolism ; Liver ; metabolism ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Murine hepatitis virus ; Potassium Channels ; genetics ; metabolism

OBJECTIVEStudies have shown that potassium channel plays a pivotal role in T cell activation. The expression of potassium channel gene KCTD9 was evidenced being highly upregulated in patients with severe hepatitis B (SHB). To understand this phenomenon further, tissue and cellular expression profiles of KCTD9 were investigated in patients with SHB.

METHODSA rabbit peptide polyclonal antibody was prepared. Various samples including peripheral blood mononuclear cells (PBMCs); livers from patients with SHB or mild chronic hepatitis B, were examined for KCTD9 expression by quantitative real time PCR and immunohistochemistry staining (IHC). Confocal microscopy was used to illustrate the localizations of the expressions.

RESULTSIncreased expression of KCTD9 was observed in PBMC in over 35.7% of the patients with SHB when compared with that of patients with mild chronic hepatitis B. In all patients, the relative value of increased KCTD9 mRNA was positively correlated with alanine aminotransferase, aspartate aminotransferase, total bilirubin and direct bilirubin but negatively with serum albumin. The expression was mainly located in hepatocytes, bile duct epithelial cells, Kupffer cells and inflammatory cells, and in the cytoplasm of PBMCs from the healthy individuals and patients with mild chronic hepatitis B, whereas in both cytoplasm and nuclei in those from patients with SHB.

CONCLUSIONThe increased expression of potassium channel gene KCTD9 correlates with disease severity in patients with viral hepatitis B.

Adult ; Female ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Middle Aged ; Monocytes ; metabolism ; Potassium Channels ; genetics ; metabolism ; RNA, Messenger ; genetics ; Young Adult

BACKGROUNDThe association between increased serum uric acid (SUA) levels and cardiovascular risk has been debated for decades. Several large studies have provided conflicting results regarding the clinical significance of elevated SUA levels in cardiovascular disease (CVD) or cerebrovascular disease. The aim of this study was to investigate the relationship between SUA and CVD and all-cause mortality and their potential diagnostic value.

METHODSA total of 3570 in-patients ranging in age from 56 to 95 years (mean (67.36 +/- 11.36) years) were selected from 20 hospitals in Beijing and Shanghai. A carefully designed questionnaire was used to gather baseline data of each patient. All patients were divided into two main groups according to their SUA levels: high SUA and normal SUA groups. Serum indices and other important parameters were measured.

RESULTSCompared with normal SUA group, high SUA group had significant difference in systolic blood pressure (SBP), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), body mass index (BMI), and age (P < 0.05 or P < 0.01). High SUA prevailed in female and patients with history of essential hypertension, while history of smoking and diabetes showed no significant difference between two groups. All-cause and CVD mortality occurred more frequently in high SUA group than in normal SUA group. In the accumulative survival analysis, high SUA group had lower survival rate than normal SUA group both in CVD and all-cause mortality. COX regression analysis indicated that the history of smoking, age and high SUA were independent risk factors for the development of CVD.

CONCLUSIONSThese preliminary observations suggest that patients with high SUA levels would face higher risk of mortality. SUA measurement may be applied as a routine predictor for clinical assessment.

Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Cardiovascular Diseases ; blood ; etiology ; mortality ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Uric Acid ; blood