蜜环菌对高血糖患者血清影响分析

蜜环菌是重要的药用真菌,具有很好的人体调节功能。研究蜜环菌对高血糖患者血清的影响,给出3个实验组,A组注射生理盐水0.3 mL.d^-1,B组注射香菇多糖10 mL·kg^-1·d^-1,C组注射蜜环菌多糖10 mL·kg^-1·d^-1,连续实验两周,测定3组高血糖患者的肿瘤细胞增长率,以及高血糖患者血清CA199水平变化情况,并对检测方法的准确率进行验证分析。实验结果表明,注射蜜环菌多糖的患者体内肿瘤细胞增长率较低,蜜环菌多糖能够降低高血糖患者血清CA199水平,与其他2组对比,具有明显的统计学意义,且血清CA199检测方法准确率较高。表明蜜环菌能提高人体免疫能力。     

菌核侧耳β-葡萄糖苷酶液体培养条件的优化

通过单因素和正交试验,对菌核侧耳产β-葡萄糖苷酶液体培养条件进行优化。结果表明:以黄豆浆为氮源,葡萄糖为碳源,初始pH为6.0产酶条件最优。培养温度为25℃、装液量50mL/250mL、摇瓶转速为150r/min培养72h后,酶活力达最高,为1.86U/g(湿菌丝体)。     

金针菇液体菌种发酵罐内深层培养条件的研究

研究了金针菇908液体菌种在发酵罐内深层培养过程的生长变化及影响条件。结果显示，培养过程中适宜的温度为24-25℃，pH5．5～7．0，接种量15％。培养前期24h，通气比1vvm，搅拌器转速180r／min；在24～60h适宜的通气比为1．2vvm，搅拌器转速210r／min。24-60h为菌丝的对数生长期。60h时的菌球数为720个／mL，还原糖含量5．96g／100mL。氨基氮含量9．54mg／100mL，纤维素酶、淀粉酶、漆酶及蛋白酶的酶活分别为1．85u、2．38u、0．092u及0．184u。     

香菇菌柄基部多糖的提取及其清除DPPH自由基作用的研究

以香菇菌柄基部和纤维素酶为试材,研究了纤维素酶解法提取香菇菌柄基部多糖的最佳工艺.结果表明:粉碎粒度200目,酶解温度35℃,酶解时间3h,香菇菌柄基部多糖的提取率可达到5.124％;初步纯化后的多糖对DPPH自由基有较强的清除活性,当清除率为50％时,香菇多糖的IC50为0.13 mg/mL.     

白灵菇多糖对力竭运动大鼠抗疲劳及抗氧化作用探析

为探究白灵菇多糖对力竭运动大鼠抗疲劳和抗氧化作用,选取250只Wistar雄性大鼠,采用随机数字表法平均分成5组,使其进行为期30 d跑台运动,观察白灵菇多糖对大鼠抗氧化能力的影响及对大鼠NO-NOS、ATPase活性的影响。在使用白灵菇多糖后,大鼠机体的谷胱甘肽过氧化物酶(GSH-Px)活力、SOD活力与-SH含量明显升高,丙二醛(MDA)水平显著降低;休息12 h后,大鼠机体的一氧化氮合酶(NOS)能力、一氧化氮(NO)水平与运动前水平一致,因此,白灵菇多糖能够增强机体抗氧化能力,并且能够增强机体的恢复能力。     

香菇的发酵研究及相关检测

经研究,筛选所获香菇菌株能适应液体培养环境,48h前为其调整期,48～144h处于增殖生长期;发酵培养96～120h,生物量即达30g/1OOmL(鲜重);提制的香菇营养液,氨基酸含量达1851,49mg/1OOmL,多糖含量为14.75mg/100mL;提制的香菇多糖粉是一种甘露糖肽,行较强的免疫活性,得率在0.3％左右。     

茯苓液体培养条件研究及其营养浓缩液的相关检测

研究表明，液体培养条件下，茯苓经0～48h的适应期，48～144h的增殖期后进入稳定期，发酵液pH骤降。经检测，发酵液中含有包括十六烷酸和十八烷酸在内的有机酸；发酵罐培养120h，生物量≥30g/l00mL(鲜重)。茯苓营养浓缩液多糖含量≥5．0mg/mL，氨基酸含量≥480mg/100mL。动物实验证明，茯苓营养浓缩液具有显著的抗疲劳和提高机体免疫力作用，其重金属和微生物指标均符合国家标准。     

蛹虫草多糖对小鼠抗疲劳作用的研究

采用雄性ICR小鼠随机分组,分别灌胃蒸馏水、不同剂量的蛹虫草(Cordyceps militaris)多糖(低、中、高剂量组分别每天灌胃75、150、450mg/kg BW),40d后建立疲劳模型,比较力竭游泳时间、肝肌糖原水平、乳酸含量和机体相关的乳酸脱氢酶和超氧化物歧化酶活力。结果表明:高剂量蛹虫草多糖能够显著延长小鼠力竭游泳时间,中高剂量多糖显著提高小鼠肝肌糖原水平,有效的遏制运动后小鼠乳酸生成量,而低剂量多糖显著增加运动后小鼠乳酸脱氢酶活力,3个剂量的多糖均能提高运动后小鼠血清中超氧化物歧化酶的活力。     

黄芪多糖在草菇冷藏保鲜中的应用

分析4℃处理24、48 h的草菇（Volvariella volvacea）渗出液代谢组；用200μmol·L-1黄芪多糖溶液浸泡草菇后在4℃放置24、48 h，测定草菇β-葡萄糖苷酶活性和转录组，检测糖基转移酶基因VVO＿02307表达量。结果表明：4℃处理24、48 h草菇渗出液代谢组分明显不同；与4℃处理24 h草菇比较，处理48 h的草菇中蔗糖含量较低，亚油酸含量较高。与对照比较，黄芪多糖溶液浸泡的草菇在4℃处理24 h时自溶不明显，β-葡萄糖苷酶活性显著降低。转录组分析结果显示，黄芪多糖增强草菇对低温胁迫的耐受性，可能是通过提高N-聚糖生物合成通路活力实现的。研究结果可为草菇低温保鲜技术研发提供参考。     

灵芝原生质体制备与再生条件的研究

本文研究了不同酶浓度、酶解温度、酶解时间、渗透压稳定剂及菌龄等对灵芝菌丝原生质体制备与再生的影响，结果表明，菌龄为60h的菌丝，采用浓度为2％的溶壁酶以0．6M的甘露醇作渗透压稳定剂于pH6.5，30℃下酶解2h，当酶解液中湿菌体含量对0．1g/mL时，其原生质体数可达2^*10^7个/mL，且再生率达8．5％。     

Mechanisms underlying the protection of berberine against liver injury induced by lipopolysaccharide in mice

AIM: To investigate the protective effects of berberine against liver injury induced by lipopolysaccharide in mice and the mechanisms underlying its protective effect. METHODS: The male mice were divided randomly into control, berberine group, LPS group and berberine treatment group. Mice were administered intragastrically with distilled water (0.01 mL/g) or 5 g/L neutral sulfate berberine (0.01 mL/g) once a day for 5 days and injected intraperitoneally with normal saline or LPS （0.02 mL/g，28 mg/kg）at 1 h after gavage on day 5. Blood was collected for determining alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, the content of tumor necrosis factors-α (TNF-α) at 10 h and 2 h after LPS or normal saline injection, respectively. Furthermore, the liver tissue was processed, and histological changes and ultrastructure in liver were observed with light and electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in liver were also detected. RESULTS: Both ALT and AST activities in serum in LPS group were higher than those in control and berberine treatment group. LPS increased the serum TNF-ɑ content at 2 h after injection, which was reversed by berberine pretreatment. The histological examination showed that LPS caused severe hepatic cell edema, degeneration, apoptosis and even necrosis, and ultrastructure observation demonstrated that LPS induced mitochondrial swelling, condensation and margination of chromatin, irregular nuclear envelope in hepatocytes. The above pathological changes produced by LPS were attenuated by berberine pretreatment. Moreover, MDA contents in liver tissue were higher in LPS group than control and berberine treatment group, but there were no significant difference in SOD activity between berberine treatment and LPS group. CONCLUSION: Berberine has a protective effect on LPS-induced liver injury in mice, the mechanisms may be related to its decreasing the production of TNF-α, inhibiting lipid peroxidation and protecting mitochondria.     

Effect of clinorotation on in vitro cultured explants of Mentha piperita L

An in vitro culture system was used to study the influence of gravity on axillary shoot formation and adventitious root regeneration in Mentha piperita L. The direction of the gravity vector was altered by displacing stem node explants in different orientations. Also, microgravity conditions were simulated by rotating the explants on a horizontal clinostat so that the main axis of nodes was either parallel (Cpa) or perpendicular to the clinostat axis (Ccp and Ccf, centripetally and centrifugally oriented, respectively). Mint nodes were cultured on solidified Linsmaier and Skoog's medium [Physiol. Plant. 18 (1965) 100] adding a filter-sterilized aqueous solution of 2 mg/l benzyladenine (BA) in half of the cultures. The proliferation of axillary shoots as well as adventitious root formation were not affected by altering upright explant orientation. On the contrary clinorotation was able to modify plantlet development. In absence of BA, leaf width was hindered by Cpa treatment and penultimate internode length was enhanced by Ccp. Furthermore, a negative effect of Cpa treatment was observed in root length parameter, while Ccp increased the root number both in absence and in presence of BA. An effect strictly connected to clinorotation in presence of BA was the occurrence of hyperhydricity. Moreover, explants under clinorotation treatments switched their gravitropic response modifying shoot curvature.     

Expression of MMP-2 and NF-κB in myocardium of mice with viral myocarditis

AIM: To observe the effects of TNF-α/nuclear factor-κB(NF-κB)/matrix metalloproteinase-2(MMP-2) pathway on the expression of MMP-2 in the mice with viral myocarditis. METHODS: Six-week-old inbred male mice were randomly assigned to control and myocarditis group. The mice in myocarditis group and control group were intraperitoneally inoculated with 0.1 mL 10^-5.69 TCID50/mL coxsackievirus B3 and vehicle (PBS), respectively. Ten mice were sacrificed at the 4th and 10th days after injection. The blood and heart specimens were harvested. The serum content of TNF-α was measured by ELISA. The myocardial levels of MMP-2, NF-κB p65 and IκBα were determined by Western blot. RESULTS: Compared with control group, the protein expression of MMP-2 and NF-κB p65 in the myocardium and the serum content of TNF-α were significantly increased in myocarditis group (P<0.05). The protein expression of IκBα was lower in myocarditis group than that in control group (P<0.05).CONCLUSION: TNF-α, NF-κB p65 and MMP-2 were higher in the mice with acute viral myocarditis. The increased expression of them might be involved in the pathogenesis of viral myocarditis.     

姬松茸发酵液对小鼠抗运动疲劳及肝组织抗氧化能力的影响

研究姬松茸(Agaricus blazei)发酵液灌胃小鼠,对小鼠抗运动疲劳及肝组织抗氧化能力的影响.试验结果表明,与对照组(10 mg/kg·d ddH2O)相比,姬松茸发酵液中剂量组(20mg/kg·d)、高剂量组(30 mg/kg·d)小鼠的负重游泳、爬杆、耐缺氧时间均显著延长,血清尿素氮和血乳酸含量显著降低,肝糖原储存量显著升高,肝组织中SOD、GSH-Px活性显著提高,MDA值显著下降.说明灌胃姬松茸发酵液,能提高小鼠抗运动疲劳功能和加强肝组织抗氧化能力.     

Relationship between polymorphism of angiotensin I converting enzyme gene insertion/deletion and ACE,PAI-1 activity in patients with myocardial infarction

AIM:To explore the relationship between polymorphism of angiotensin I converting enzyme gene insertion/deletion (I/D) and ACE, PAI-1 activity in patients with myocardial infarction (MI). METHODS:Ninety-three patients with MI and eighty-seven healthy controls were tested. ACE genomic DNA was amplified using the polymerase chain reaction (PCR). Serum ACE activity was measured by colorimetry, plasma level of PAI-1 activity was determined by spectrophotometric assay. RESULTS:① The frequency of ACE DD genotype and D alleles (32.3% and 54.3%) in MI group was significantly higher than those in control group (12.6% and 37.4%, P<0.01, respectively). ② The ACE activity in serum (216.00±58.26)U/L and plasma PAI-1 activity (0.85±0.19)AU/mL in MI group were significantly higher than those in control group (170.19±48.99)U/L, (0.66±0.20)AU/mL, P<0.01, respectively. The serum ACE activity was positively correlated with plasma PAI-1 activity both in MI group and control group (r=0.7108 and r=0.7829;P<0.01, respectively). ③ In MI group, the serum ACE activity and plasma PAI-1 activity showed a significantly higher level in subjects with DD genotype (251.64±57.76)U/L, (0.96±0.16)AU/mL than those with ID (211.47±51.87)U/L, (0.82±0.18) AU/mL and Ⅱ genotypes (179.84±52.65)U/L, (0.71±0.17)AU/mL. The serum ACE activity and plasma PAI-1 activity were significantly higher in subjects with ID genotype than those with II genotype (P<0.05). In control group, the serum ACE activity and plasma PAI-1 activity showed a significantly higher level in subjects with DD genotype (195.53±54.76)U/L, (0.78±0.20)AU/mL than the subjects with Ⅱ genotype (154.98±52.74)U/L, (0.59±0.17)AU/mL (P<0.05). CONCLUSION:The increased ACE activity caused by DD polymorphism may play an important role in elevating the level of plasma PAI-1. The DD genotype of ACE is associated with high PAI-1 level. The genetic variation of ACE contributes to the balance of fibrinolytic pathway, indicating the pathogenesis mechanisms linking to the ACE I/D genotype and MI.     

草菇子实体提取物的抗氧化活性

为考察草菇(Volvariella volvacea)子实体提取物(VBS)的抗氧化活性,测定了对两种自由基的清除率及其总还原力,结果表明:对羟自由基和DPPH自由基的清除率以及总还原力均与VBS的浓度存在正相关的量效关系,IC50值分别为1.40、0.67、0.63mg/mL。以ICR雄性小鼠为研究对象,灌胃给药12d,再用CCl4花生油溶液灌胃,8h后分别测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙氨酸氨基转移酶(ALT)和谷草转氨酶(AST)的活力及丙二醛(MDA)的含量,各项指标在血清中的结果表明,VBS中剂量组(200 mg/kg)的抗氧化活性较好,而在肝脏中则表明VBS高剂量组(400mg/kg)的抗氧化活性较好。     

Effect of Qingyi decoction on p-STAT3 expression in pancreas of mice with severe acute pancreatitis induced by L-arginine

AIM: To explore the role of STAT3 signaling pathway in acute pancreatitis induced by L-arginine, and the mechanisms of Qingyi decoction(QYD) treatment on severe acute pancreatitis (SAP). METHODS: The Kunming mice were randomly divided into 3 groups (n=10): control group, SAP group and QYD treatment group (SAP+QYD). The mice in SAP group and SAP+QYD group were intraperitoneally injected with 20% L-arginine (3 g/kg, bid). The mice in SAP+QYD group were also administered intragastrically with QYD(10 mL/kg) 30 min after the second injection of 20% L-arginine and twice a day for the following 2 days. The mice were anesthetized and sacrificed 72 h after SAP induction. The activity of amylase was measured in serum, the relative pancreatic weight was assayed, and the activity of myeloperoxidase (MPO) was analyzed to evaluate the neutrophil infiltration in lung tissues. The morphology of pancreas and lung was observed. The protein of pancreas was extracted to detect the expression of p-STAT3 by Western blotting. The mRNA expression of monocyte chemoattractant protein-1(MCP-1) was determined by real-time PCR. RESULTS: Compared with control group at 72 h, L-arginine induced SAP with increased serum amylase activity, pancreatic wet weight ratio and MPO activity in lung tissues (P<0.05). In SAP+QYD group, the activity of amylase, pancreatic wet weight ratio and MPO levels were significantly lower than those in SAP group (P<0.01). Compared with control group at 72 h, the pancreas and lung were obvious injured, the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas tissues increased significantly in SAP group. Compared with SAP group, the pathologic damage of the pancreas and lung tissues, the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas were significantly reduced in SAP+QYD group. CONCLUSION: The expression of p-STAT3 in pancreas increases in the mice with SAP induced by L-arginine. The activation of STAT3 may take part in the development of SAP. Inhibition of STAT3 activation in pancreas is one of the mechanisms of QYD treatment for SAP.     

A mouse model of upper respiratory tract mucosal immunity dysfunction induced by cold stimulation

AIM: To establish a mouse model of upper respiratory tract mucosal immunity dysfunction induced by cold stimulation. METHODS: Mice were stimulated with cold by placing the animals in-20 ℃ environment for 5 min, 10 min, 15 min or 20 min. The secretory immunoglobulin A(SIgA) level and lysozyme activity in the mouse saliva were determined for qualification. RESULTS: The mortality rate was 50% in the group of cold stimulation for 20 min. Compared with the control mice, the SIgA level and lysozyme activity significantly decreased in the group of cold stimulation for 15 min (P<0.05 and P<0.01,respectively). Decreased lysozyme activity was only observed in the group of cold stimulation for 10 min (P<0.05). The data in the group of cold stimulation for 5 min were almost similar to those in control group. CONCLUSION: A mouse model of upper respiratory tract mucosal immunity dysfunction was successfully established by cold stimulation.     

Mechanisms for a decrease in mortality of LPS-challenged mice by rhynchophylline

AIM: To observe effect of rhynchophylline （Rhy） on mortality and organ injury in endotoxemic mice and further investigate the mechanisms of its actions. METHODS: Male mice were randomly assigned into control, LPS, Rhy +LPS and Rhy group, and injected subcutaneously with normal saline (0.05 mL/10 g), or rhynchophylline once a day for 3 d, 1 h after subcutaneously treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally. Survival rate was recorded every 12 h for 6 d. In another experiment, 12 h after LPS injection, the left lung and intestine tissue sections were prepared for histological analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D),the serum was collected to detect the concentrations of alanine aminotransferase（ALT）, aspartate aminotransferase (AST ), bloodureanitrogen (BUN) and creatinine (Cr). In addition, the concentrations of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and interleukin-10 (IL-10) in serum at 2 h after LPS challenge were detected by enzyme-linked immunosorbent assay. The concentration of NO in serum at 8 h was detected by enzymic method. The effect of Rhy on survival rate of mice subjected to cecal ligation and puncture (CLP) was also observed. RESULTS: Mortality of mice challenged with LPS alone was higher significantly than that in control at 24 h after LPS challenge, pretreated with Rhy at a dose of 8 or 16 mg/kg increased markedly the survival rate of LPS-challenged mice. However, Rhy at a dose of 8 mg/kg significantly increased mortality of mice subjected to CLP. In the histological analysis, severe inflammation was observed both in the lung and intestine tissues in the LPS group. LPS elevated lung W/D, the levels of ALT, AST, BUN, Cr, TNF-α, IL-1β, IL-10 and NO in serum. Pretreatment with Rhy had no obvious improvement in the lung and intestine tissue injury, and no significant depression in the lung W/D and the serum levels of ALT, AST, BUN, Cr, IL-1β, IL-10 and NO, but decreased the level of TNF-α in serum significantly in LPS -treated mice. CONCLUSION: Pretreatment with Rhy reduces the mortality in endotoxemic mice, but not decrease the mortality of mice challenged with CLP, at least in part, through inhibiting the synthesis and secretion of TNF-α.