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AIM:To observe the effect of B-HT933, a selective α2-adrenoceptor agonist, on lipopolysaccha-ride ( LPS )-induced TNF-αproduction in neonatal rat cardiomyocytes and to explore the underlying mechanisms . METHODS:The neonatal rat cardiomyocytes were cultured .The localization of α2A-adrenoceptor in the cardiomyocytes was examined by immunofluorescence staining .The cardiomyocytes were exposed to LPS or/and B-HT933 for different time.The level of TNF-αin the supernatants and the mRNA expression of TNF-αwere detected by ELISA and real-time PCR, respectively.In addition, LPS-associated signal molecules in the cardiomyocytes were also examined by Western blotting.RESULTS: Immunofluorescence staining showed that α2A-adrenoceptors were localized in the cardiomyocytes . LPS stimulated TNF-αproduction in the cardiomyocytes in a dose and time-dependent manner .B-HT933 pretreatment sig-nificantly inhibited the expression of TNF-αat mRNA and protein levels in LPS-treated cardiomyocytes .Furthermore, LPS exposure induced IκBαand p38 phosphorylation in cardiomyocytes and only IκBαphosphorylation was prevented by B-HT933 treatment.CONCLUSION:α2A-adrenoceptors are present in neonatal rat cardiomyocytes and its agonist B -HT933 inhibits LPS-induced TNF-αproduction in cardiomyocytes via suppressing IκBαphosphorylation .
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BACKGROUND: CpG oligodeoxynucleotide (ODN) is a type of highly effective immune adjuvant with low toxicity, which has an extensive application in gene therapy for many diseases. However, the specificity for species and cells leading to low uptake by cells and degradation by nuclease blocks its clinical application. OBJECTIVE: To explore the specific delivery and its immunologic efficacy of CpG ODN targeting B lymphocytes of umbilical cord blood by CD40 ligand-receptor-mediated carrier system. DESIGN, TIME AND SETTING: An observation and control experiment was performed at the Department of Hematology, and Department of Pediatric, the First Affiliated Hospital of Jinan University from April 2004 to October 2007. MATERIALS: Fresh umbilical cord blood with heparin was obtained from healthy, natal infant. Informed consent was obtained from his parents, and the experiment was approved by the hospital Ethics Committee. METHODS: CD40 ligand (CD40L)-EDC-PLL-CpG ODN conjugated complex was prepared. Mononuclear cells (MNCs) from umbilical cord blood were co-cultured with conjugated complexes. Uptake rate, mean fluorescence intensity of FAM marked CpG ODN, expressions of MNCs, proliferations of lymphocytes and the IgG levels of culture supematants were detected by flow cytometry, fluorescence techniques, MTT assay and ELISA, respectively. MAIN OUTCOME MEASURES: The uptake rate, the mean fluorescence intensity of CpG ODN by MNCs, subgroups and proliferations of lymphocytes, and IgG levels of culture supematants. RESULTS: Compared to the pure CpG ODN group, the uptake rate of the conjugated complexes group was higher (98%), the peak level of up-taking occurred earlier, and intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, and CD20+ was increased, A value and IgG levels in supematants were all higher than that of the control group. CONCLUSION: CD40 tigand-receptor-mediated carrier system is helpful for CpG ODN delivery targeting to B lymphocyte, enhancing its immunological efficiency.
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Objective To investigate the effects of siRNAs targeted protein kinase C ?(PKC?) on the proliferation and apoptosis of A549 cell line.Methods Six PKC? siRNAs were designed and chemical synthesized. Candidate PKC? siRNA was transfected into A549 cells,PKC? mRNA level was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Then,the effects of more effective PKC? siRNAs on A549 were further investigated by using clone formation assay,Hoechst 33258 staining,propidium iodide (PI) staining,Annexin V-FITC and PI dual staining and western blot. Results Six candidate PKC? siRNAs were designed and named as No.1,No.2,No.3,No.4,No.5 and No.6 PKC? siRNA. Compared with controls,PKC? mRNA levels were all significantly downregulated and the cell proliferation was inhibited in A549 cells treated with candidate PKC? siRNAs except No.5 (P
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Traditional teaching mode of biochemistry experiment couldn’t satisfy the demand of cultivating creative medical talents in new era. Offering designing experiment is the developing direction of experiment teaching reforms. But in the practice,there are various of obstacles which prevent the reforming process.
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AIM: To investigate effects of Retinervus luffae fructus (RLF) on level of serum lipid and body weight in hyperlipidemia rats. METHODS: Thirty-two male SD rats were randomly divided into four groups: control group (A), hyperlipidemia group (B), hyperlipidemia + RLF group (C), RLF group (D). Both group A and C were fed normal diet every day, while group B and group D fed high fat diet. Meanwhile, group C and D were administered with RLF solution at the dose of 10 mL/kg, respectively for 14 days, while group A and B were administered with drinking water. RESULTS: (1) At the end of experiment, a significant reduction was found in the levels of serum total cholesterol (TC) and triglyceride (TG) of group C animals treated with RLF solution; (2) The levels of serum TC of group D was progressively decreased compared to the level of serum TC at the beginning of experiment; (3) The level of high-density lipoprotein cholesterol (HDL-C) of group C remained unaltered 8d after treatment with RLF solution; (4) The body weight in group C was obviously lower than that in group B. CONCLUSION: RLF had an obvious hypolipidemic effect on hyperlipidemia rats. It can inhibit the decrease in the HDL-C and the increase of body weight in rats. [
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AIM: To assess the antiradiation effect of oligomeric procyanidins. METHODS: The hemolysis, malondialdehyde(MDA) levels of mouse liver homogenates and the broken degree of DNA single-strain after being irradiated by ultraviolet were examined. RESULTS: Oligomeric procyanidins significantly reduced the hemolysis of erythrocytes induced by ultraviolet irradiation, inhibited the increase in MDA levels of irradiated mouse liver homogenates ( P
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Objective To formulate genuine case mix plans using medical record information available in China. Methods An account was given of the research background of case mix studies, the current status of the studies and applications at home and abroad, the case mix model for measuring medical "output", data sources and statistical methods of classification. Results Case mix plans for army inpatients, civilian inpatients and outpatients were respectively formulated, and software for calculating the case mix index of a hospital which can run on Windows 95/98 computer systems was developed. Conclusion The theoretical, methodological and data conditions necessary for the formulation of case mix plans based on the first pages of medical records in China are now ripe and the case mix plans formulated on the basis of data from large samples can now be used for the control of medical costs, the assessment of a hospital's medical "output" and the estimation of the financial compensations for hospitals.
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98%), the peak level of uptake occurred earlier, intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, CD20+ increased significantly. A_~570 values of MNCs proliferation and IgG levels in supernatant were all higher. CONCLUSION: CD40 ligand-PLL carrier system may delivery CpG ODN targeting to B lymphocytes, enhancing its immunological efficiency.
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AIM: To examine the microsatellite instability (MSI) and loss of heterozygosity (LOH) in head and neck squamous cell carcinomas (HNSCC). METHODS: 36 cases of HNSCC were analyzed with 15 microsatellite markers from chromosome 3,5,6,8,9,13,17 and 18. RESULTS: Among the 36 cases of HNSCC, 27.8%(10/36)of samples showed MSI in one to eight microsatellite markers. High frequent MSI occurred at D17S520(22.9%), D6S105(16 7%)and D8S264(13 9%). LOH was detected on the site of 9p21-p22 and 3p14. No correlations were found between allelic instability and grade or stage of the tumor. CONCLUSION: Our data suggest that MSI is a common genetic change in HNSCC. Tumor suppressor genes related to HNSCC may harbor at chromosome 9p21-p22 and 3p14 regions. [
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AIM: To design,prepare and screen out functional small interfering RNA(siRNA) for specifically silencing proliferating cell nuclear antigen(PCNA) gene expression and effectively inhibiting cell proliferation on human hepatoma cell line SMMC-7721,human gastric carcinoma cell line SGC-7901 and human colorectal carcinoma cell line Caco2.METHODS: PCNA siRNA was designed based on previous studies about design guidelines and synthesized in vitro by T7 Mega short script reaction kit according to the manufacturer's instructions.After purification,the integrities of siRNA were identified through 19% denaturing polyacrylamide gel electrophoresis,and the concentrations of the generated siRNA were measured by testing the absorbance at 260 nm using a spectrophotometer.Four synthesized double-strand siRNA were transfected into three types of carcinoma cell lines by LipofectamineTM2000 reagent,respectively.WST-8 assay was employed to examine the proliferative inhibitions of these three cell lines.The subsequent alterations on PCNA mRNA and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochemistry,respectively.RESULTS: 3 sequences,No.2,No.3 and No.4 PCNA siRNA showed effective inhibitions on tumor cells among the 4 candidate siRNA,and a single dose of 50 nmol/L of No.2,No.4 PCNA siRNA showed the most effective inhibitory rates as more than 62% at 48 h after transfection.50 nmol/L of No.2 and No.4 PCNA siRNA transfection caused 72% decrease of PCNA mRNA level and almost completely loss of the protein in human Caco2 cells.CONCLUSION: No.2 and No.4 PCNA siRNA have been screened out in this study,which show the capability to effectively down-regulated PCNA expression and significantly inhibit the proliferation of carcinoma cells.The optimal concentration is 50 nmol/L and satisfactory effects are achieved 48 h after transfection in vitro.
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AIM: To investigate the effect of tea polyphenols (TP) on alcohol-induced liver injury in mice.METHODS: 40% alcohol(0 5 mL) and TP (5 mg) were administered intragastrically, the liver MDA level and serum alanine transaminase (ALT) activity were determined. The effect of TP on MDA level in alcohol-treated liver in vitro was also examined.RESULTS: Pretreatment with TP significantly inhibited alcohol-induced liver MDA increase in mice in vivo and in vitro , the increase in serum ALT induced by alcohol was also reduced by pretreatment with TP (0 5 mg). TP at a dose of 5 mg, administered 1 h after alcohol treatment, also suppressed increase in liver MDA level stimulated by alcohol. CONCLUSION: These results demonstrated that TP has a protective effect on alcohol-induced liver injury in mice.
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AIM: To further investigate the role of central corticotropin-releasing hormone in stress-induced hyperthermia and lipopolysaccharide (LPS)-induced fever in the rat. METHODS: Test substances were administered intracerebroventricularly (icv) via a third ventricle cannula. Body temperature responses were monitored at 30 min intervals using colonic thermistor probes. Arginine vasopressin (AVP) level in the ventral septal area (VSA) determined by radioimmunoassay. RESULTS: In normal saline controls, rats were handled to take the colonic temperature, their body temperature significantly increased with a peak of (0.88?0.31)℃. The injection (icv) of ?-helical CRH(9-41), a CRH-41 receptor antagonists, markedly attenuated the stress-induced hyperthermia within 90 min after injection of normal saline. LPS(300 ng, icv) stimulated a biphasic rise in the colonic temperature, the 3.5 h thermal response index (TRI 3.5 ) and AVP levels in the VSA of LPS-treated rats were higher than those of control rats. The AVP responses to LPS were inhibited significantly by blockade of central CRH actions using ?-helical CRH(9-41) (5 ?g ,icv) administered 10 min prior to LPS, while ?-helical CRH(9-41) (5 ?g ,icv) resulted in exacerbated febrile responses to LPS(300 ng, icv). CONCLUSION: Central CRH plays an important role in stress-induced hyperthermia. The injection (icv) of ?-helical CRH(9-41) enhances markedly LPS-induced fever in rats. CRH is a dual action molecule in LPS-induced fever, which itself mediates LPS-induced fever, at the same time, and limits the rise in body temperature during fever through actions of AVP in the VSA and glucocoticoids.