Objective: To investigate anti-bacterial, anti-biofilm and hemolytic characteristics of synthetic linear peptides based on the structure of lipopeptide 6-2. Methods: A series of five linear peptides (LP1-LP5) was synthesized through a solid-phase synthesis based on the structure of lipopeptide 6-2, which is produced by the Bacillus amyloliquefaciens anti-CA strain. The structures of these peptides were identified by MS and H-1-NMR. Results: The water solubility of the five new compounds was improved and the hemolytic properties were significantly reduced compared with those of lipopeptide 6-2 after the introduction of water-soluble groups, such as AEEA and glucose. The anti-bacterial activity of LP5 was effectively maintained. LP5 was selected to evaluate the inhibition effect upon the biofilm formation. Conclusion: LP5 exerted an identical inhibitory effect upon the biofilms as lipopeptide 6-2.

Leucine aminopeptidase 3 is involved in the progression and metastasis of several cancers. This study aimed to screen anti-tumor lead compounds targeting leucine aminopeptidase 3. The compounds' suppression effect on enzyme activity and anti-tumor activity were evaluated through a series of assays. Leucine aminopeptidase 3 overexpression K562 cells were used as an enzyme source to screen 43 natural marine compounds. Compounds 5 and 6 exhibited high suppression effect on leucine aminopeptidase 3 activity. Cell activity tests indicated that both compounds have an anti-proliferative effect on triple-negative breast cancer cells. Wound healing assay and transwell invasion assay showed that both compounds could inhibit the migration and invasion of breast cancer cells. Immunoblot analysis exhibited that both compounds could downregulate the expression of metastasis-related proteins fascin and matrix metalloproteinase-2/9. A molecular dynamic simulation process was applied to discover the key features of compounds 5 and 6 in binding to leucine aminopeptidase 3 active site. This study described the anti-tumor effects of two leucine aminopeptidase 3 small molecule inhibitors. Taken together, compounds 5 and 6 could be used as anti-tumor lead compounds targeting leucine aminopeptidase 3.

The expression of leucine aminopeptidase 3 (LAP3) is associated with the prognosis for and malignant transformation of many types of tumors. Therefore, a LAP3 inhibitor may represent a new strategy for cancer therapy. Evaluating the suppression of enzyme activity by an LAP3 inhibitor is essential. Right now, leucine aminopeptidases (LAPs) purified from the porcine kidneys are the only enzymes that can be used to evaluate the suppression of enzyme activity by an LAP3 inhibitor. This approach cannot accurately reflect the suppression of human LAP3 by an inhibitor. The current study developed a new method with which to evaluate the suppression of enzyme activity by an LAP3 inhibitor. Total protein from K562 cells seldom catalyzed the LAP3 substrate. A lentivirus was used to induce K562 cells to overexpress LAP3 (K562-LAP3). After puromycin screening, flow cytometry data indicated that 98.8% of cells expressed green fluorescent protein. The expression of LAP3 in K562-LAP3 cells was also assessed using Western blotting. K562-LAP3 cells were lysed with ultrasonication. Total protein was used as an enzyme source and L-leucine p-nitroaniline hydrochloride was used as a substrate to measure enzyme activity. Total protein from K562-LAP3 cells catalyzed the substrate more than that from K562 cells did. The LAP3 inhibitor ubenimex was used as a positive control to evaluate the suppression of LAP3 enzyme activity. Results indicated that ubenimex significantly inhibited the enzyme activity of LAP3. This approach provides a convenient and accurate way to evaluate the suppression of enzyme activity by an LAP3 inhibitor.

临床药理学研究人体与药物相互作用及其规律,是药学专业研究生的主干课程之一.随着人才需求日益更新,课程教学过程中的问题更加凸显.对临床药理学教学内容、教学方法、教学结果评价等三个方面进行分析,为提高教学质量提供参考.

Glucagon-like peptide-1 (GLP-1) is a peptide with multiple functions in regulating blood glucose with the mechnism still not throughly understand. AMP-activated protein kinase (AMPK) plays an important role in glucose and energy homeostasis, especially in type 2 diabetes mellitus (T2DM) pathophysiology. This study explores GLP-1 analogue exendin-4 (Ex-4) regulating glucose balance by activating AMPK and regulation gluconeogenesis in insulin-resistant cell model (IR/ HepG2). IR/HepG2 cells were treated with Ex-4, AMPK activator or Ex-4 pretreated with AMPK inhibitor Compound C. The change of cell morphology, glucose consumption and lipid content in IR/HepG2 cells were examined. The expression of AMPK and p-AMPK in IR/HepG2 treated by Ex-4 were determined by Western blot and immunochemisthistology (IHC) assay. Key enzymes of glucose metabolism, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G-6-Pase) in IR/HepG2 cells treated by Ex-4 were detected by Western blot, immunohistochemistry and qRT-PCR. The results showed that Ex-4 treatment increased the expression of p-AMPK and reduced the expression of G-6-Pase and PEPCK in IR/HepG2 cells. Pretreatment with AMPK inhibitor Compound C counteracts the effect of Ex-4. These findings suggest that Ex-4 fulfilled it glucose regulation effect by phosphorylation of AMPK and decreased the expression of enzymes involving gluconeogenesis.

Astragalus polysaccharide (APS) is a natural compound extracted from astragalus membranaceus which has an antidepressant activity. Initially APS was studied for its immunomodulatory potential, and later was found to exhibit multiple pharmacological effects, including anti-inflammatory activity. More recently APS was shown to attenuate the lipopolysaccharide (LPS) induced neuroinflammation and improve the learning and memory ability of rats. The major objectives of this study were to investigate whether APS would exhibit antidepressant effects in an animal model of depression induced by LPS, and whether this effect might be associated with regulating nuclear factor-kappaB (NF-kappa B) and mitogen-activated protein kinase (MAPK) signaling pathways. Three groups of Wistar rats were injected LPS (i.p.), two groups of which were pretreated with APS (200 mg or 400 mg, i.p.). Behaviors were evaluated by forced swim test, saccharin preference test and open field test. Levels of NF-kappa B p65, phospho-NF-kappa B p65, phospho-I kappa B alpha, ERK1/2, JNK, p38 MAPK, phospho-ERK1/2, phospho-JNK and phospho-p38 MAPK in hippocampus and hypothalamus were measured to assess the activities of NF-kappa B and MAPK signaling pathways. In addition, levels of TNF-alpha, IL-1 beta and IL-6 (both protein and mRNA levels) in hippocampus and hypothalamus were determined. Results showed LPS induced depressive behaviors, as well as activated the NF-kappa B and MAPK signaling pathways in rats. APS treatment dose-dependently alleviated depressive-like symptoms and inhibited the activation of NF-kappa B and MAPK signaling pathways induced by LPS. The data indicate an antidepressant-like activity of APS in a LPS-induced animal model of depression possibly via inhibition of NF-kappa B and MAPK signaling pathways.

Objective:To investigate the effects of Exendin-4 (Ex-4) on the expressions of lipid metabolism related genes in the human liver cancer HepG2 cells with insulin resistance (IR),and to elucidate the effect of Ex-4in improvement of IR.Methods:The HepG2 cells in logarithmic growth phase were induced into IR model with high concentration of insulin,then divided into control group (HepG2 cells),IR group (HepG2 cells were treated with insulin,HepG2-IR cells),and Ex-4 group (HepG2-IR cells were treated with Ex-4).Glucose oxidase (GOD-POD)kit was used to detect the consumption of glucose.The cell morphology and intracellular lipid drip formation were observed by Oil red O staining.The triglyceride (TG) level in cells was detected by kit;qRT-PCR was used to detect the mRNA expression levels of acetyl-CoA carboxylase (ACC),fatty acid synthase (FAS),sterol regulatory element-binding protein-1c (SREBP-1c) and apolipoprotein B100 (apoB100).Results:Compared with control group (HepG2 cells),the glucose consumption in the HepG2-IR cells in IR group was significantly decreased (P＜0.01).Compared with IR group,the glucose consumption in the HepG2-IR cells in Ex-4 group was increased (P＜0.05).The Oil O red staining results showed that compared with control group,the fat percentage in the HepG2-IR cells in IR group was increased (P＜0.05);compared with IR group,the fat percentage in Ex-4 group was decreased (P＜0.05).Compared with control group,the level of TG in the cells in IR group was significantly increased (P＜0.01);compared with IR group,the level of TG in the cells in Ex-4 group was significantly decreased (P＜0.05).The qT-PCR results showed that compared with control group,the expression levels of ACC FAS and SREBP-1cmRNA in the cells in IR group were increased (P＜0.01),and the expression level of apoB100 mRNA was decreased (P＜0.05);compared with IR group,the expression levels of ACC,FAS and SREBP-1c mRNA in the cells in Ex-4 group were decreased (P＜0.05),and the expression level of apoB100 mRNA was increased (P＜0.01).Conclusion:Ex-4 can regulate the expressions of lipid metabolism related genes in the HepG2 cells and improve IR.

Aminopeptidase N (APN, also known as CD13) is involved in cellular processes of various types of tumors and a potential anti-cancer therapeutic target. Here, we report the effect of an APN inhibitor 4cc in enhancing sensitivity of hepatocellular carcinoma (HCC) cell lines and xenograft model in response to 5-fluorouracil (5-FU) in vivo and in vitro. The treatment of the combination of 4cc with 5-FU, compared to the combination of bestain with 5-FU, markedly suppressed cell growth and induced apoptosis of HCC cells, accompanying the increase in the level of reactive oxygen species (ROS) and followed by a decrease in the mitochondrial membrane potential (ΔΨM). Furthermore, the combination of 4cc and 5-FU showed a significant inhibitory effect on the growth of HCC xenograft tumors. In addition, following the treatment of 4cc, APN activity and clonogenic formation and the number of CD13-positive cells in PLC/PRF/5 cells were significantly decreased, suggesting that 4cc may also inhibit liver cancer stem cells by CD13 inhibition. These results showed that the APN inhibitor 4cc synergizes antitumor effects of 5-FU on human liver cancer cells via ROS-mediated drug resistance inhibition and concurrent activation of the mitochondrial pathways of apoptosis.

Objective To study the sleep improvement function of DHA +Lecithin.Methods By measur-ing the direct sleep time ,the sleep time that induced by pentobarbital sodium ,the sleep incidence rate of sudbissociative dose and the latency time that induced by barbital sodium ,it could observe the mice sleep improvement function .Results Each dose group had no significant difference on the mice weight and direct inducing sleep .DHA could shorten the la-tency time that induced by barbital sodium (P<0.05).DHA+Lecithin could increase the sleep incidence rate of sud-bissociative dose(P<0.05) and shorten the latency time that induced by barbital sodium (P<0.05).Conclusion DHA+Lecithin can improve the mice sleep time .

The fast and diversified development of modern society requires medical technology talents to grasp the international cutting-edge information and the latest development,most of which come from english reports,so the importance of english teaching in medical colleges and universities is increasingly apparent.The teaching and research room of pharmacology of our university made a series of exploration in bilingual teaching reform by strengthening the integration of bilingual teaching and english case teaching and have made some achievements in the training of highly skilled bilingual medical technology talents.

Pharmacology classes has been given with full English to medical international students since year 2005 in our school.According to the different characteristics,we improved and explored the teaching methods of pharmacology gradually.In this paper,in order to improve the teaching level and teaching effectiveness of pharmacology,we described some teaching experience and methods,and gave the comparison results before and after the teaching methods changing.

Objective To study the effect of Shenmai injection on prevention of oxaliplatin chemotherapy induced toxicity.Methods Sixty patients were randomly divided into two groups,30 cases in each group.Patients in control group were treated with oxaliplatin 100 mg/m2;patients in experimental group were given 100 mg/m2 of oxaliplatin with Shenmai injection 80 mL/d in 5% dextrose intravenous drip concurrently,each case was completed 4 cycles of chemotherapy after participating in the evaluation.Results Compared with control group,the immune function of patients in experimental group enhanced and the neurotoxicity of oxaliplatin reduced.Conclusion Shenmai injection can reduce oxaliplatin chemotherapy induced toxicity.

OBJECTIVE To observe the effects of Astragalus polysaccharides (APS) on the BG, insulin and C-peptide in serum, ultrastructure and Fas expression of pancreatic beta-cells in diabetes mellitus (DM) rats. METHODS Thirty DM rats induced by streptozotocin (STZ) were randomly divided into three groups: DM group, APS 200 mg/kg group, APS 400 mg/kg group, another 10 normal rats were taken as the control group. The drug was given by intraperitoneal for 6 weeks. The level of BG was determined by ONE TOUCH II machine. The levels of insulin and C-peptide in serum were measured by radioimmunoassay. The expression of Fas was observed by immunohistochemistry. Moreover, the TEM was used to observe the ultrastructure of pancreatic beta-cells. RESULTS (1) DM rats showed significant increase in BG compared with control group (P < 0.05). APS could decrease the level of FG (P < 0.05). (2) DM rats showed significant decrease in insulin and C-peptide in serum compared with control group (P < 0.05). APS has no appearance effects on the levels of them (P > 0.05). (3) The expression of Fas of beta-cells was significantly increased in DM rats, which was significantly inhibited by APS treatment. (4) It showed degenerative changes of pancreatic beta-cells ultramicroscopic structure of the DM rats, while APS treatment could significantly improve the damage. CONCLUSION APS exerts its therapeutic effects on DM, which maybe related to the significant decreasing of the Fas expression and inhibiting the apoptosis of beta-cells.

目的:观察养阴清热解毒汤对干燥综合征(SS)外周血T细胞亚群变化的影响.方法:20例患者给予养阴清热解毒汤治疗,疗程4周.采集治疗组患者治疗前后的外周血以及10例健康对照组外周血检测T细胞亚群变化.结果:干燥综合征患者与健康对照组比较外周血CD4+T淋巴细胞水平明显减低,CD8+T淋巴细胞水平明显升高,CD4+/CD8+减低,差异有统计学意义(P＜0.05).使用养阴清热解毒汤治疗后CD4+T淋巴细胞水平较治疗前升高,CD8+T淋巴细胞水平较治疗前降低,CD4+/CD8+较治疗前升高,治疗前后比较差异有统计学意义(P＜0.05).结论:T细胞亚群分布紊乱可能在干燥综合征发病中起重要作用,养阴清热解毒汤可纠正干燥综合征患者此紊乱.

To improve effectiveness of pharmacology teaching,PBL(problem-based learning) method was used as a new education concept in post-graduate students.The students were divided into two groups randomly,PBL education concept was used in teaching of the experimental group students,a traditional concept of LBL(lecture-based learning) was used in the control group.The evaluation results between two groups were different.It was concluded that PBL education concept is superior to the traditional method LBL in pharmacology learning for post-graduate students,PBL is a concept worthy of promotion in teaching of post-graduate students.

Objective:To investigate the effect of JNKK1 expression with potential metastasis and prognosis of breast carcinoma. Methods:Immunohistochemistry was used to detect and analyze JNKK1 expression in 103 cases of breast carcinoma. Results:The positive expression rate of JNKK1 was 44.7%(46/103) ,and the negative rate was 55.3%(57/103) . Breast carcinoma patients with positive expression of JNKK1 had higher metastatic rate(P0.01) and poorer survival rate in 5 years(P 0.01) . Conclusion:The expression of JNKK1 is closely related to the potential of metastasis in breast carcinoma. JNKK1 may be a prognostic and treatment indicator in breast carcinoma.

Aim: To observe the effects of astragalus polysaccharides (APS) on the ultrastructure and Fas expression of pancreatic beta-cells in diabetes mellitus (DM) rats. Methods: Thirty DM rats induced by streptozotocin (STZ) were randomly divided into three groups: DM group, APS 200 mg·kg -1·d-1 group, APS 400 mg·kg -1·d-1 group, another 10 normal rats were taken as the control group. The drug was given by intraperitoneal for 6 weeks. The protein expression of Fas was observed by immunohistochemistry. The TEM was used to observe the ultrastructure of pancreatic beta-cells. Results: Circled digit one The expression of Fas of beta-cells was significantly increased in DM rats, which was significantly inhibited by APS treatment. Circled digit two It showed degenerative changes in the ultramicroscopic structure of pancreatic beta-cells of DM rats, while APS treatment could significantly improve the damage. Conclusion APS exerts its therapeutic effects on DM, which may be related to the significant decrease of the Fas expression and inhibiting the apoptosis of beta-cells.

正常角膜组织透明,无血管,周围血管终止于角膜缘,形成血管网,营养成分由此扩散入角膜.无血管是角膜的主要特征,也是维持角膜透明的重要条件.在病理状态下,新生毛细血管由角膜缘处侵入角膜内,称为角膜新生血管(corneal neovascularization,CNV).内皮抑素(endostatin)是一种新型内源性新生血管生成抑制因子,可特异性抑制新生血管内皮细胞的增殖,封闭新生血管的形成,在肿瘤实验过程中表现出极强的抑瘤活性.实验表明,内皮抑素对多种起源的新生血管内皮细胞增殖有抑制作用,而不影响静止的血管内皮细胞,内皮抑素具有明显抑制角膜新生血管生长的作用,而无毒副作用,为临床防治CNV提供了一种新的途径.

Aim To probe into the mechanism of the therapeutic effect of astragalus polysaccharides(APS)against diabetes mellitus rats by measuring renal medullary aquaporin-2(AQP-2),the change of urinary volume in 24 hs,as well as observing kindey ultramicroscopic structures.Methods Thirty diabetes mellitus(DM)rats induced by streptozotocin(STZ)were randomly divided into three groups:DM group,APS 200 mg·kg-1·d-1 group,and APS high dosage group.Another 10 normal rats were taken as the control group.The drug was given by introperitonial injection for 6 weeks.A 24 h urine collection through simple metabolic cage was made to measure urinary volume;the RT-PCR technique was used to determine the levels of AQP-2 mRNA expression in kidney medulla.Kidney medulla ultramicroscopic structures were observed by TEM.Results Diabetic rats showed a significant increase in urinary volume compared with control rats.The urine amount of low and high dosage APS treatment could obviously increase the urinary volume(P0.05);the expression of AQP-2 mRNA was up-regulated in the kidney of diabetic rats.Low and high dosage APS treatment could alleviate the over expression of AQP-2 mRNA(P0.05).It showed degenerative changes of distal tubule and collecting tubes chief cells ultramicroscopic structures of the DM,APS low dose and high dose treatment group in contrast with the control group,while APS Long-term treatment could significantly improved the damage.Conclusion APS exerts its therapeutic effects on diabetes mellitus,which may be related to the significantly decreased expression of AQP-2 mRNA in the kidney medullary.

Aim: To probe the mechanism of the therapeutic effect of APS against DM rats by measuring the level of TNF-α, as well as observing the change of caspase-3 protein in pancreatic beta cells. Methods: Thirty DM rats induced by STZ were randomly divided into three groups: DM group, APS low dosage group, APS high dosage group, and another 10 normal rats were taken as the control group. The drug was given for 6 weeks. The research included measuring the level of TNF-α in serum and the expression of caspase-3 protein in pancreatic beta cells. Results: Circled digit one The level of TNF-α was higher in model group than that of the normal group (P < 0.05). APS could decrease the level of TNF-α (P < 0.05); Circled digit two The expression of caspase-3 protein of beta cells in islet was significantly increased in DM rats, which was significantly inhibited by APS treatment. Conclusions: APS can decrease the level of TNF-α and inhibit the apoptosis of beta cells in islet, which may be one of the mechanisms of APS improving DM.