关于灵芝改善睡眠功能的研究

通过直接睡眠实验、延长戊巴比妥钠睡眠时间、提高戊巴比妥钠阈下催眠剂量实验和缩短巴比妥钠睡眠潜伏期实验,观察灵芝提取物对睡眠的作用。结果:经口给予小鼠不同剂量的灵芝提取物30天,对照组及3个剂量组在给予受试物60分钟内,均未发现有直接睡眠效应。与对照组比较,灵芝提取物在45.0mL/kg.bw剂量组时能延长戊巴比妥钠诱导的小鼠睡眠时间(P<0.01)、提高戊巴比妥钠阈下剂量小鼠入睡动物发生率(P<0.05)、缩短巴比妥钠睡眠潜伏期(P<0.05),即灵芝提取物具有改善睡眠的功能。灵芝提取物对小鼠体重增长无影响。     

灵芝提取物及复合制剂改善睡眠和免疫调节的研究

目的：研究灵芝提取物及复合制剂改善睡眠和免疫调节的作用;方法：小鼠给予灵芝提取物及复合制剂30d后,通过直接睡眠试验、延长戊巴比妥钠睡眠时间、戊巴比妥钠阈下催眠剂量试验和缩短巴比妥钠睡眠潜伏期试验评价改善睡眠作用;通过DNFB诱导的小鼠迟发性变态反应（DTH）和碳廓清试验观察免疫调节的作用;结果：与对照组相比,灵芝提取物及复合制剂能显著延长戊巴比妥钠睡眠时间、缩短巴比妥钠睡眠潜伏期（P〈0．05或P〈0．01）和增加入睡动物发生率;灵芝提取物能显著增强小鼠迟发性变态反应、碳廓清能力和免疫脏器指数（P〈0．05）;复合制剂能显著增强小鼠迟发性变态反应（P〈0．05）,对小鼠碳廓清能力和免疫脏器指数无显著影响。结论：灵芝提取物及复合制剂对小鼠睡眠和免疫功能有一定的改善作用。     

蜜环菌提取物改善睡眠及免疫调节研究

通过戊巴比妥钠睡眠试验和免疫作用试验(迟发型超敏反应和碳粒廓清),研究蜜环菌提取物对试验小鼠睡眠改善影响和免疫调节作用。睡眠试验结果表明蜜环菌提取物,对增加戊巴比妥钠诱导睡眠时间有正相关作用,小鼠入睡率也有明显提升,还可以巴比妥钠睡眠潜伏期有较为明显的降低,说明蜜环菌提取物对于改善小鼠睡眠起到了积极作用。免疫试验结果表明蜜环菌提取物可以加大迟发型超敏反应,在碳粒廓清试验中,同样可以让小鼠关键免疫细胞参数上升,可以说明蜜环菌提取物也提高了小鼠的免疫调节作用。     

猴头菌丝体对中枢神经系统的作用

深层发酵培养小刺猴头菌球提取物对小鼠自发活动具有剂量依赖性抑制作用,在给药后半小时使小鼠抑制更深,与氯丙嗪合用其作用增强,且可延长戊巴比妥钠睡眠时间,能对抗苯丙胺对中枢神经的兴奋作用。     

3种杀菌剂对葡萄白腐病的防治药效试验

采用20％戊菌唑水乳剂、10％世高水分散颗粒剂、25％势克悬浮剂等三种药剂对葡萄白腐病进行了田间药效试验,结果表明,3种药剂对葡萄白腐病均有较好的防治效果,极显著优于对照药剂70％甲基托布津,且对葡萄均无药害作用,具有良好的安全性.     

四种杀菌剂对胶孢状炭疽菌的毒力测定

以胶孢状炭疽菌为试验菌,采用生长速率法,测定了75%肟菌酯·戊唑醇水分散粒剂、60%唑醚·代森联水分散粒剂、30%戊唑·多菌灵悬浮剂、80%代森锰锌可湿性粉剂等4种杀菌剂对来自梨、葡萄和无花果等3种寄主胶孢状炭疽菌的毒力。结果表明:4种杀菌剂对胶孢状炭疽菌的毒力存在差异。肟菌酯·戊唑醇、唑醚·代森联和戊唑·多菌灵等3种复配的毒力高于代森锰锌,其中戊唑·多菌灵对3种寄主胶孢状炭疽菌的毒力最高。戊唑·多菌灵和肟菌酯·戊唑醇对3种寄主胶孢状炭疽菌的EC50比较相似。唑醚·代森联对葡萄胶孢状炭疽菌的毒力较高,其次为梨胶孢状炭疽菌和无花果胶孢状炭疽菌。     

25%氰戊·乐果可湿性粉剂防治辣椒蚜虫田间试验

通过开展25%氰戊·乐果可湿性粉剂防治辣椒蚜虫药效试验,明确了该产品防治辣椒蚜虫的田间药效、最佳使用剂量及安全性,为该产品大面积推向市场提供依据。该试验对蔬菜蚜虫的防治具有借鉴意义。     

土壤处理剂在直播芹菜田间除草效果试验

为明确33%二甲戊乐灵乳油、48%氟乐灵乳油、48%地乐胺乳油和50%扑草净可湿性粉剂作为土壤处理剂在芹菜田间的除草效果,2009年在直播芹菜田间进行试验。结果表明:48%氟乐灵乳油和48%地乐胺乳油对芹菜的出苗有影响;33%二甲戊乐灵乳油对阔叶杂草的防除效果差;除草效果最好的为50%扑草净可湿性粉剂,建议使用浓度为3 750 g/hm2。     

不同药剂处理对核桃举肢蛾的防治试验

核桃举肢蛾是天水地区主要虫害之一,通过不同防治方法试验,总结出了采取土壤深翻、树盘铺盖地膜、树盘撒施毒土等无公害防治措施的基础上,夏季喷施20%氰戊·马拉松乳油1 200倍液、20%高氯·马乳油1 200倍液、25%高效氯氟氰菊酯微乳剂2 000倍液,防治效果最佳。     

黑柄炭角菌发酵物安全性评价初探

黑柄炭角菌(Xylaria nignipes)是一种名贵的食药用菌, 生于地下1～3m废弃的白蚁巢中,产量极少,价格昂贵。我们采用生物工程技术,从天然样品中分离出菌株,实现了工厂化生产黑柄炭角菌菌丝体,并成功地开发以菌丝体为主要原料的保健品。经现代分析仪器测定,菌丝体含有丰富的蛋白质,必需氨基酸配比合理,并含有较高的钙、镁、铁、锌等矿物质与微量元素、甘露醇和蛋白多糖等活性物质。经动物试验和人群食用表明,本品具有调节内分泌代谢,增强机体免疫功能,促进血红蛋白、白细胞和红血球的生成、健脑、护脑,调节青年女子经期紊乱和改善老年人排泄功能,促进睡眠等保健作用,是一种具开发利用潜质的保健品原料。本文对黑柄炭角菌的发酵物的安全性作了研究。     

Study on the electrophysiological characteristics of ion channels of stem cell derived cardiomyocytes of mouse

AIM: To study the electrophysiological characteristics of ion channels of stem cell derived cardiomyocytes(SCDC) of mouse. METHODS: Embryonic stem cells of D₃ line(ES-D₃) were cultured on the MEF feeder layer with BRL conditioned medium, and fetal mouse heart cells(FMHC)were cultured in vitro. Then ES-D₃ cells were induced to differentiate into many kinds of cells. SCDC were harvested on day 12 after differentiation initiating and identified by electro-microscope and immunocytochemistry. SCDC and FMHC were prepared for the patch-clamp research. Sodium and calcium currents together were elicited and compared between SCDC and FMHC. RESULTS: The current characteristics of sodium and calcium channels of SCDC were very similar to FMHC. CONCLUSION: The functional expression of ion channels occurred during ES-D₃ cells differentiation and the electrophysiological characteristics of sodium and calcium channels of SCDC are very similar to FMHC.     

miRNA-122 promotes mouse embryonic stem cell-derived hepatic precursor cells to differentiate into hepatocytes

AIM：To investigate whether miRNA-122 (miR-122) promotes the differentiation of mouse embryonic stem cell (ESC)-derived hepatic precursor cells (HPCs) into hepatocytes. METHODS：Mouse ESCs were initially induced to differentiate into HPCs by stimulating with fibroblast growth factor 4 (FGF-4), sodium butyrate and dexamethasone (Dex) sequentially. Then a recombinant adenovirus expressing vector pAV.Ex1d-CMV>miR-122/IRES/eGFP was constructed by Gateway technology and transfected into the mouse ESC-derived HPCs 9 d after induction, so as to gain the cells with stable high expression of miR-122. The morphological changes of transfected cells were observed under inverted phase-contrast microscope. The liver-specific gene expression levels were detected by real-time RT-PCR. The liver-specific protein expression levels were also detected by immunofluorescence method. The liver functions were assessed by indocyanine green (ICG) uptake experiment, glycogen staining and urea synthesis function test. RESULTS：The mouse ESCs were successfully induced into HPCs by stimulating with FGF-4, sodium butyrate and Dex sequentially. At 6 d after transfection of miR-122, the morphology of the cells was closer to the mature hepatocytes. The mRNA levels of liver-specific genes such as albumin (ALB), transthyretion, α1-antitrypsin, glucose-6-phosphatase, cytokeration 8, cholesterol 7α-hydroxylase and cytochrome P450 3A4 were up-regulated. The expression levels of liver-specific proteins such as ALB and cytokeratin 18 were increased, while alpha-fetoprotein was decreased. The results of ICG uptake experiment, glycogen staining and urea synthesis function test indicated that the hepatocyte functions were strengthened as compared with control group. CONCLUSION：The combination of FGF-4, sodium butyrate and Dex successfully induces the mouse ESCs into HPCs. Over-expression of miR-122 effectively promotes the differentiation and maturation of mouse HPCs.     

In vitro differentiation of exocrine pancreatic cells from mouse embryonic stem cells

AIM: To explore the effects of sodium butyrate, activin A and dexamthasone on inducing mouse embryonic stem (ES) cells to differentiate into exocrine pancreatic cells in vitro. METHODS: E14 mouse ES cells were cultured in suspension to form embryonic bodies (EBs). The EBs were cultured with differentiating medium containing different concentrations of sodium butyrate, and the spontaneously differentiated ES cells were used as control. Exocrine pancreatic genes such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR at different time points to determine the optimal concentration and exposure time of sodium butyrate. Furthermore, activin A or dexamthasone was also used to explore the effects on exocrine differentiation. After that, the combination of sodium butyrate, activin A and dexamthasone was used to promote the differentiation of exocrine pancreatic cells from ES cells. During the differentiation course, the gene expressions of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR. Morphological changes were investigated by phase contrast microscopy. Amylase expression was examined by immunofluorescence staining. RESULTS: Exocrine pancreatic gene expressions such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected in spontaneously differentiated EBs. A relatively lower concentration of sodium butyrate with a shorter exposure time significantly promoted those above gene expressions as compared to that of spontaneously differentiated EBs. Activin A and dexamethasone induced upregulation of exocrine gene expression. The combination of activin A, sodium butyrate and dexamethasone significantly enhanced the mRNA levels of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase. Under the treatment of activin A, sodium butyrate and dexamethasone, differentiated cells were polygonal in shape with large, round, and center-situated nuclei. According to the observation of immunofluorescence staining, amylase was positive expressed at the final stage. CONCLUSION: These data indicate that exocrine pancreatic differentiation of ES cells is induced by sodium butyrate, activin A and dexamethasone. The combination of pancreatic inducing factors improves the differentiating efficiency.     

Sodium butyrate induces mouse embryonic stem cells to differentiate into hepatocytes in vitro

AIM:To explore mechanism by which sodium butyrate induces mouse embryonic stem cells (ES) to differentiate into hepatocytes in vitro. METHODS:E14 mouse ES cells were cultivated in a routine way, and then cultivated in suspension to form embryonic bodies (EBs). EBs were transferred into 6-well culture dishes and 3 mmol/L sodium butyrate was added into the culture medium. Morphological changes were investigated by phase contrast microscopy. α-fetoprotein (AFP), albumin (ALB) and cytokeratin 18 (CK18) were examined by immunofluorescence staining. AFP, ALB, α1-antitrypsin (AAT) and TTR mRNA were assayed by RT-PCR. Proportion of ALB positive cells was analyzed by flow cytometry. Periodic acid Schiff (PAS) reaction and indocyanine green (ICG) uptake assay were performed to assess the characteristic hepatocyte function of the differentiated cells. RESULTS:In the presence of sodium butyrate, parts of ES cells differentiated into a population with epithelial morphology similar to mouse hepatocytes. AFP and TTR mRNA expression were observed at 7 d, and ALB and AAT mRNA expressed at 14 d. Hepatocytes specific markers, ALB, AFP and CK18 were positive expression in immunofluorescence staining at 14 d. PAS reaction and ICG uptake were positive for the hepatocyte-like cells. CONCLUSION:Mouse ES cells can be induced into hepatocyte-like cells by sodium butyrate efficiently, and these ES cells-derived hepatocytes possess characteristic hepatocytic function.     

Pathological changes and activity of SOD and CAT in the damaged retina induced by sodium iodate in rats

AIM: To investigate the pathological changes and the activity of superoxide dismutase(SOD) and catalase(CAT) in the damaged retina induced by sodium iodate in rats.METHODS: The SD rats were randomly divided into 4 groups, 40 mg/kg, 50 mg/kg and 60 mg/kg of sodium iodate treatment groups and control group. Rats were sacrificed on day 1, 4, 7, 14 after the injection of sodium iodate. The right eyeballs were immediately enucleated and fixed. Routine pathological examination was carried out. And the retinal tissues were extracted from the left eyeballs. The activity of SOD and CAT were measured by photometry. RESULTS: The pathological examination showed that 40 mg/kg and 50 mg/kg of sodium iodate didnt induce damage of retina on day 1, while 60 mg/kg of sodium iodate induced the damage of retinal pigment epithelium and disarrangement of outer nuclear layer compared with control group. Each dosage on day 4,7,14 induced more significant damage than that on day 1, with waveform damage changes and decreasing depth of outer nuclear layer (P<0.05 or P<0.01). As the photometry test showed, compared with control group, 60 mg/kg of sodium iodate induced significant decrease of SOD and CAT activity on day 1 and day 4 (P<0.05); each dosage induced significant decrease of SOD and CAT activity on day 7 and day 14 (P<0.05 or P<0.01). CONCLUSION: This suggests that all of 40 mg/kg, 50 mg/kg and 60 mg/kg of sodium iodate could induce damage of retinal pigment epithelium and outer nuclear layer, and they could also break down the antioxidative system. Sodium iodate could have the dose-dependent effect on the onset time and the level of retinal damage.     

Inhibitory effects of genistein on mouse allergic contact dermatitis

AIM: To investigate the inhibitory effects of genistein (Gen) on mouse allergic contact dermatitis (ACD). METHODS: The animal model of ACD was induced by DNFB. The effects of different doses of Gen on mouse ear swelling，body weight, histopathological changes in mouse ear skin, thymus index and spleen index were observed. RESULTS: All groups of Gen inhibited mouse ear swelling induced by DNFB significantly. The infiltration of inflammatory cells and thymus index were also reduced. However, the increase in mouse body weight was not affected. Low dose of Gen increased spleen index, high dose of Gen decreased spleen index. CONCLUSION: Genistein has significant inhibitory effects on mouse ACD induced by DNFB.     

Effects of N-acetylcysteine on the lipopolysaccharide-induced MAPK phosphorylation in mouse liver

AIM: To investigate the effects of antioxidant N-acetylcysteine (NAC) on the lipopolysaccharide (LPS)-induced MAPK phosphorylation in mouse liver. METHODS: 54 male mice were divided into three groups: control (n=6), 0.9% sodium chloride 0.2 mL ip; LPS group (n=24): LPS 5 mg ip; NAC+LPS group (n=24): NAC 150 mg·kg-1·d-1 ip, for 3 d; LPS 5 mg ip after 1 h of NAC administration at 3rd day. The liver was excised with carbrital anesthesia after LPS or 0.9 % sodium chloride injection at 0.5 h, 1 h, 2 h and 6 h for GSH and MDA assays. The protein extracted from liver was assayed for the phosphorylation of MEK1/2, ERK1/2, p38 MAPK by Western blotting. TNF-α in liver was assayed by radioimmunoassay. RESULTS: MDA in the liver was decreased remarkably and the GSH in the liver was increased significantly by NAC pretreatment. The phosphorylation of MEK1/2, ERK1/2 and p38 MAPK in liver were inhibited significantly by NAC pretreatment after LPS challenge. Meanwhile, TNF-α in liver was decreased markedly. CONCLUSION: Reactive oxygen species plays a critical role in MAPK signaling during the LPS induced acute liver injury. NAC partially inhibits LPS-induced MAPK signaling by antioxidant effect and decreases TNF-α production.     

Effects of oxidized low density lipoprotein and lysophosphatidylcholine on cholesterol efflux from mouse peritoneal macrophage foam cells

AIM:To explore the effects of oxidized low density lipoprotein (OxLDL) and one of its component— lysophosphatidylcholine (LPC) on cholesterol efflux from mouse macrophage foam cells.METHODS:(1) Cholesterol efflux induced by apoAI from mouse peritoneal macrophage foam cells loaded with OxLDL or acylated LDL(AcLDL) was measured. (2) Cholesterol efflux induced by LPC and apoAI from macrophage foam cells separated from normal or apoE gene deficient (E⁰) mouse loaded with AcLDL were measured.RESULTS:(1) When the macrophage foam cells were incubated with apoAI, cholesterol efflux from AcLDL-induced macrophage foam cells increased significantly compared to that of OxLDL-induced macrophage foam cells. (2) LPC promoted cholesterol efflux from macrophage foam cells in relation to both dosage and time. When LPC was incubated with E⁰ mouse macrophage foam cells, the released cholesterol mass was significantly lower than that of normal mouse macrophage foam cells. It was also found that cholesterol efflux induced by apoAI normally occurred in E⁰ mouse macrophage foam cells.CONCLUSIONS:(1) OxLDL accumulated cholesterol in macrophages and impair cholesterol efflux. (2) LPC induced cholesterol efflux from macrophage foam cells, which may occur via apoE pathway.     

Roles of novel cannabinoid chemicals O-1602 and cannabidiol in DSS-induced mouse colitis

AIM:To investigate the roles of O-1602 and cannabidiol(CBD) in dextran sulfate sodium(DSS)-induced mouse colitis. METHODS:The model of colitis was induced in C57BL/6 mice by drinking water containing 4% DSS for 7 days. The model mice were treated with O-1602(5 mg/kg), CBD(1 mg/kg) or SB203580. A colitis scoring system was used to evaluate the colon local lesion, and the systemic inflammatory responses were observed by detecting the plasma levels of tumor necrosis factor α(TNF-α), interleukin 6(IL-6) and cytokine-induced neutrophil chemoattractant 1(CINC-1), and the activity of myeloperoxidase(MPO) in the lung tissues. The expression of G protein-coupled receptor 55(GPR55) was detected by the method of immunohistochemistry. The expression of p38 and phosphorylated p38(p-p38) in colon tissues was determined by Western blotting. RESULTS:O-1602 and CBD improved the pathological changes in the mice with DSS-induced colitis and decreased the plasma levels of TNF-α, IL-6 and CINC-1, and the activity of MPO in the lung tissues(P<0.05). Lower expression of p-p38 was observed after treatment with O-1602, CBD and SB203580(P<0.05). The expression of GPR55 was mainly in the submucosa of mouse colon tissues. CONCLUSION:O-1602 and CBD show protective effect on the mice with experimental colitis, and the anti-inflammatory roles of O-1602 and CBD are related to the inhibition of p38 MAPK. The expression level of GPR55 in the submucosa of mouse colon tissue is low.