粗状假丝酵母(Candida valida T2)生产脂肪酶的发酵条件

对粗状假丝酵母产生脂肪酶的培养条件进行了研究。结果表明,该菌株产脂肪酶的适宜培养基组成为:桐油15mL/L,黄豆粉30g/L,糊精10g/L,硝酸铵10g/L,MgSO4·7H2O1g/L,K2HPO42g/L,Tween800.5g/L;最佳培养为温度30℃、发酵液起始pH6,摇瓶发酵脂肪酶活力达到1467U/mL。     

产脂肪酶菌Geotrichum candidum NS3的固定化及其稳定性

从南昌地区含油土样中,筛选到一株脂肪酶产生菌白地霉(Geotrichum candidum NS3).菌株摇瓶发酵的水解酶活为321 U/g干细胞,合成酶活为0.54 U/g干细胞.以聚氨酯泡沫为载体对菌株Geotrichum candidum NS3进行固定化培养和稳定性研究,结果显示,聚氨酯泡沫颗粒尺寸6 mm×6 mm×6 mm,密度27 kg/m3,摇瓶培养60 h,有73.8%的细胞进入聚氨酯泡沫中生长固定,载体固定细胞干重达到2.32 g/g载体.电镜图片显示白地霉(Geotrichum candidumNS3)在载体孔隙内和脊壁上缠绕充盈,生长良好,固定结构稳定.固定化细胞颗粒连续5批次催化反应,相对水解酶活保持率和固定细胞干重保持率分别达到63.1%和71%,具有良好的细胞固定稳定性和酶活保持率.     

产过氧化氢酶菌株培养条件的优化

通过对溶壁微球菌培养基优化,确定最佳发酵培养基,并对发酵工艺条件进行了初步的优化.最适碳源是麦芽糖,最适氮源是蛋白胨和酵母膏,最近无机盐是MnSO4;该菌株产酶的最佳培养基配方为:60 g麦芽糖,13 g蛋白胨,8 g酵母膏,1 g MnSO4,5 g NaCl,2 g NaH2 PO4·2H2O,2 g K2HPO4·3H2O,0.5 g MgSO4·7H2O,0.5 g CaCl2,1 000 mL水.从该菌株发酵过程曲线发现,该菌株大量产酶期在12-14 h.摇瓶发酵最适初始pH值为7.0,250 mL三角瓶装液量为25mL,接种体积分数为5%,培养温度为37℃.     

出芽短梗霉产色素能力弱化菌株的筛选

出芽短梗霉在发酵过程中会产生一种与黑色素相似的黑色物质 ,并且这种物质会牢固地粘附在短梗霉多糖上 .对出芽短梗霉Aureobasidium pullulanB 1菌株进行6 0 Co诱变 ,获得了一株产色素能力缺失型菌株Co3,其菌落和发酵液颜色为白色或淡绿色 .红外光谱和磁核共振图谱表明该菌株的产物与标准短梗霉多糖样品有相同的结构 .     

血脂肪酶在急性重症胰腺炎早期诊断中的价值

目的:评价血脂肪酶在急性重症胰腺炎早期诊断中的价值.方法:采用酶显色法对61例急性重症胰腺炎,28例慢性胰腺炎及18例正常对照者进行血脂肪酶检测.结果:检出急性重症胰腺炎患者血脂肪酶(1 036.23±948.85) U/L,慢性胰腺炎患者血脂肪酶(276.54±543.18) U/L,正常对照组血脂肪酶(103.85±60.97) U/L,3组比较有统计学差异(P<0.0 001).急性重症胰腺炎、慢性胰腺炎和正常对照组血脂肪酶测量值不同,且重症胰腺炎患者测量值比慢性胰腺炎组及正常对照组测量值高.结论:血脂肪酶可以作为急性重症胰腺炎的诊断指标之一,具有一定的特异性.     

圆弧青霉PG37碱性脂肪酶的发酵工艺条件

研究了圆弧青霉PG3 7碱性脂肪酶的发酵工艺条件 ,优化了PG3 7的摇瓶最适产酶条件 .其中 ,发酵培养基的组成为 (g/dL) :豆饼粉 3 .0 ,玉米浆 3 .0 ,磷酸氢二钾 1 .0 ,硫酸镁 0 .1 ,大豆磷脂0 .5,柠檬酸钠 0 .0 5,花生油 0 .2 ;发酵培养基起始 pH 7.5,发酵温度 (2 9± 1 )℃ ,摇床转速 2 50r/min ,发酵周期 96h ,发酵期间于 3 6,54,72h分别流加 0 .4 g/dL花生油 .在此条件下 ,PG3 7的脂肪酶产率为 2 0 60 μmol/ (min·mL) .PG3 7在 2 5L的实验室小罐中的产酶水平为 1 90 0 μmol/ (min·mL) .     

我国报告甲型H1N1病例已逾百例世卫组织将警戒级别提至最高级严格防控甲型H1N1流感

卫生部10日通报,我国内地共新增11例甲型H1N1流感确诊病例.至此我国内地共报告111例甲型H1N1流感确诊病例,治愈出院58例.     

碱性脂肪酶的高效液相色谱和质谱分析

以常压层析系统纯化的碱性脂肪酶为出发蛋白,采用高效反相色谱对其进一步分离纯化,达到N末端氨基酸测序及肽谱分析所需的纯度.用电喷雾质谱测得脂肪酶的相对分子质量为27 217±1.对该脂肪酶的胰蛋白酶水解条件及肽段的相对分子质量等进行了研究.     

黑色素生物合成抑制剂产生菌株的筛选

采集了中国 1 1个省的土样和草样共 2 3 1份 ,分离出了 1 70 0株链霉菌 .筛选结果是 :3 9 2 4号菌株在比基尼链霉菌筛选模型中抑制圈的直径为 2 0mm ,属于直径较大者 ;其黑色素生物合成抑制剂初提物对CV 1细胞的急性细胞毒性 (致毒质量浓度 )为 1 0mg/mL ,属于毒性较小者 .根据以上试验结果 ,作者确定 3 9 2 4号菌株为黑色素生物合成抑制剂研究的试验菌株 .     

无溶剂体系酶促合成甘油中碳酸偏酯

根据中碳酸转化率的大小 ,对 5种不同来源的固定化脂肪酶筛选 ,其中固定化脂肪酶SSW的酶催化转化率最高 ,辛酸和癸酸转化率分别为 91.5 %和 93.5 % ;对脂肪酶SSW酶促合成甘油辛酸偏酯的反应条件优化 ,适宜反应条件为 :敞口反应器中反应温度 5 5℃ ,每克酸加酶量 10 0U/g ,辛酸 /甘油投料摩尔比 1∶1.1,甘油初始加水量 4 % (质量分数 ) .实验范围内中碳酸品种对脂肪酶SSW不显示基质特异性 .     

间质性膀胱炎发生前后膀胱组织组胺受体变化的动物实验研究

目的 探讨大鼠间质性膀胱炎发生前后膀胱组织中4种组胺受体(H1R、H2R、H3R和H4R)表达的变化. 方法体质量250～300 g的雌性SD大鼠30只,随机分为实验组(20只)和对照组(10只).实验组采用硫酸鱼精蛋白加氯化钾经尿道膀胱灌注建立间质性膀胱炎动物模型,2个月后处死,对照组直接处死.切取2组大鼠膀胱组织后行免疫组化染色,利用IPP 4.5图像分析软件计算各组组胺受体平均吸光度(-A)值,并进行统计学比较. 结果4种组胺受体主要表达于膀胱黏膜层,实验组H1R的(-A)值为0.054±0.031、H2R为0.032±0.021、H3R为0.047±0.033、H4R为0.149±0.191,对照组分别为0.017±0.011、0.018±0.015、0.014±0.011、0.060±0.039.实验组H1R、H2R和H3R的(-A)值明显高于对照组(P＜0.05),H4R的(-A) 值与对照组比较差异无统计学意义(P＞0.05). 结论H1R、H2R和H3R在间质性膀胱炎大鼠膀胱黏膜中表达显著升高,可能与间质性膀胱炎的发生相关,H3R可能是治疗间质性膀胱炎的新靶点.     

Histamine H(1) receptor activation inhibits the proliferation of human prostatic adenocarcinoma DU-145 cells

BACKGROUND: Histamine stimulates cell proliferation in some tumor cell lines through the activation of H(1) receptors coupled to phosphoinositide hydrolysis. We therefore set out to study the presence of H(1) receptors in the prostate cancer cell line DU-145 and the effect of their stimulation on cell growth. METHODS: The presence of histamine receptors was studied by radioligand binding. Phosphoinositide hydrolysis was assessed by measuring [(1)H]-inositol phosphate ([(1)H]-IPs) accumulation and changes in the intracellular concentration of free Ca(2+) ([Ca(2+)](i)). Proliferation was assessed by cell counting and by [(1)H]-thymidine incorporation. RESULTS: DU-145 cells express H(1) receptors (110+/-14 fmol/mg of protein) whose stimulation results in [(1)H]-IPs accumulation (602+/-23% of basal, EC(50) 2.2+/-0.4 microM) and calcium mobilization (resting level 96+/-5 nM, Delta[Ca(2+)](i) 517+/-32 nM, EC(50) 6.2+/-0.1 microM). Incubation with histamine (100 microM, 24 hr) resulted in a decrease in both cell number and [(1)H]-thymidine incorporation, blocked by the H(1) antagonist mepyramine (1 microM). CONCLUSIONS: Histamine inhibits the proliferation of DU-145 cells through the activation of H(1) receptors coupled to phosphoinositide hydrolysis.     

Effects of Propofol on H-reflex in Humans

Background: Depression of spinal cord motoneuron excitability has been proposed to contribute to surgical immobility. The H-reflex, which measures [alpha]-motoneuron excitability, is depressed by volatile anesthetics, whereas the action of propofol is unknown. The objective of this study was to determine the effects of propofol anesthesia on the H-reflex.

Methods: In 13 patients (group 1), H-reflex was measured before (T0), 3 min after (T1), and 10 min after (T2) a 2-mg/kg bolus dose of propofol, followed by an infusion of 10 mg [middle dot] kg-1 [middle dot] h-1. Ten patients (group 2) were studied when propofol was given via a programmable pump set to a propofol blood concentration of 6 [mu]g/ml, and 10 patients (group 3) were studied with the pump set to 9 [mu]g/ml. Latencies and amplitudes of H-reflexes (H0, H1, H2) and M-responses (M0, M1, M2) of the soleus muscle were recorded, and H/M ratios (H0/M0, H1/M1, H2/M2) were calculated.

Results: In group 1, H-reflex amplitudes and the H/M ratio were diminished after induction with propofol (H0vs. H1, P = 0.033; H0/M0vs. H1/M1, P = 0.042). After 10 min of propofol infusion, the H2/M2 ratio was still decreased versus H0/M0 (P = 0.031). In group 2, no difference was detected. In group 3, propofol depressed H-reflex amplitudes at T2 (H0vs. H2, P < 0.01), and amplitudes were also lower at T2 than at T1 (H1vs. H2, P < 0.01). In this group, the H/M ratio decreased from T0 to T2 (H0/M0vs. H2/M2, P < 0.002).     

Effects of propofol on H-reflex in humans

BACKGROUND: Depression of spinal cord motoneuron excitability has been proposed to contribute to surgical immobility. The H-reflex, which measures alpha-motoneuron excitability, is depressed by volatile anesthetics, whereas the action of propofol is unknown. The objective of this study was to determine the effects of propofol anesthesia on the H-reflex. METHODS: In 13 patients (group 1), H-reflex was measured before (T0), 3 min after (T1), and 10 min after (T2) a 2-mg/kg bolus dose of propofol, followed by an infusion of 10 mg x kg(-1) x h(-1). Ten patients (group 2) were studied when propofol was given via a programmable pump set to a propofol blood concentration of 6 microg/ml, and 10 patients (group 3) were studied with the pump set to 9 microg/ml. Latencies and amplitudes of H-reflexes (H0, H1, H2) and M-responses (M0, M1, M2) of the soleus muscle were recorded, and H/M ratios (H0/M0, H1/M1, H2/M2) were calculated. RESULTS: In group 1, H-reflex amplitudes and the H/M ratio were diminished after induction with propofol (H0 vs. H1, P = 0.033; H0/M0 vs. H1/M1, P = 0.042). After 10 min of propofol infusion, the H2/M2 ratio was still decreased versus H0/M0 (P = 0.031). In group 2, no difference was detected. In group 3, propofol depressed H-reflex amplitudes at T2 (H0 vs. H2, P < 0.01), and amplitudes were also lower at T2 than at T1 (H1 vs. H2, P < 0.01). In this group, the H/M ratio decreased from T0 to T2 (H0/M0 vs. H2/M2, P < 0.002). CONCLUSIONS: During steady state conditions using propofol as the sole agent, a depression of the H-reflex is observed only at a high blood concentration of 9 microg/ml. The authors suggest that immobility during propofol anesthesia is not caused by a depression of spinal motoneuron circuit excitability.     

Histone H1 vaccine therapy for overcoming acute rejection in experimental organ transplantation

OBJECTIVE: In a rat tolerogenic orthotopic liver transplantation (OLT) model, the recipient serum (post-OLT serum) shows strong immunosuppressive activity. In our previous reports, we suggested that autoreactive antibody (Ab) against histone H1 is a major immunosuppressive factor in this serum. The present study sought to determine whether up-regulation of anti-histone H1 Ab by histone H1 vaccination led to tolerance. MATERIALS AND METHODS: Using mixed lymphocyte reactions (MLR) and heterotopic heart transplantations (HHT), the alloreactive T-cell responses and allograft survivals of histone H1-immunized rats were compared with those of control rats. Cytokine and cellular profiles were determined by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. RESULTS: The alloreactive T-cell response of histone H1-immunized rats was significantly lower than that of control rats, although there was no difference in nonspecific T-cell activation between the 2 groups. The allograft survival of histone H1-immunized rats was significantly prolonged after HHT. The major histocompatibility complex (MHC) class II and CD25 molecules of histone H1-immunized rats were significantly down-regulated compared with those of control rats. Moreover, the serum cytokine profile was modified by the immunization with histone H1. CONCLUSIONS: These results suggest that histone H1 vaccination of transplant recipients leads to the production of immunosuppressive factors and the modification of cytokine/cellular profiles.     

Involvement of autoimmunity against nuclear histone H1 in liver transplantation tolerance

BACKGROUND: Our recent studies suggested that anti-histone H1 autoantibody (auto-Ab) plays an important role in experimental and clinical liver allograft tolerance as a natural immunosuppressive factor. The present study aimed to explore how the autoimmune response against histone H1 is involved in tolerance induction. METHODS: The measurement of anti-histone H1 auto-Ab and immunohistochemical analysis were performed in serum and liver allografts after orthotopic liver transplantation (OLT). To compare the auto-Ab response against histone H1 between the recipients of rejector (DA-LEW) and tolerogenic (DA-PVG) OLT models, na?ve recipients were immunized with calf thymus histone H1. The immunosuppressive state of histone H1-immunized rats was assayed by mixed lymphocyte reaction (MLR). RESULTS: Anti-histone H1 Ab titer was transiently increased during the rejection phase after OLT (days 7-21) in the DA-PVG combination, while no such response was confirmed in the DA-LEW acute rejection model. Nuclear histone H1 antigens were found in the cytoplasm and the extracellular environment in liver allografts at the rejection phase in the tolerogenic model but not in the rejector model, resulting from the transient induction of anti-histone H1 auto-Ab in recipient PVG rats after OLT. Low dose and short-term immunization with histone H1 upregulated the anti-histone H1 Ab titer in na?ve PVG rats, which exhibited a low allogeneic immune response, while no such response was found in na?ve LEW rats. CONCLUSIONS: These results suggest that the sensitivity to nuclear antigens such as histone H1 may be a key factor determining the acceptance or rejection of donor liver grafts, at least in DA-PVG and DA-LEW combinations.     

血红素加氧酶-1预处理对大鼠肺泡Ⅱ型上皮细胞氧化损伤的影响

目的 评价血红素加氧酶-1(HO-1)预处理对大鼠肺泡Ⅱ型上皮细胞氧化损伤的影响.方法 成年健康雄性SD大鼠,体重180～220 g,原代培养肺泡Ⅱ型上皮细胞,经鉴定后随机分为6组(n=8):对照组(C组),不给予任何药物,继续培养5 h;H2 O2组,加入0-5 mmol/L H2 O2,孵育3 h;不同浓度HO-1预处理组(H1-4组),分别加入0.01、0.10、1.00、10.00μmol/L HO-1孵育2 h,然后加入0.5 mmol/L H2 O2,孵育3 h.孵育结束后,于倒置相差显微镜下观察细胞形态,并进行肺泡Ⅱ型上皮细胞计数,测定细胞活力.结果 C组、H2～4组肺泡Ⅱ型上皮细胞大部分贴壁,呈圆形,胞质均匀,胞浆内含颗粒状物质;而H2 O2组和H1组肺泡Ⅱ型上皮细胞内可见反光增强的空泡,上清液中有较多的细胞碎片.与C组比较,H2O2组和H1组细胞计数和细胞活力降低(P＜0.05),H2～4组细胞计数和细胞活力差异无统计学意义(P＞0.05);与H2O2组和H1组比较,H2～4组细胞计数和细胞活力升高(P＜0.05);H2O2组和H1组间,H2～4组间细胞计数和细胞活力差异无统计学意义(P＞0.05).结论 0.10～10.00μmol/L HO-1预处理可减轻大鼠肺泡Ⅱ型上皮细胞氧化损伤.     

关节腔内注射盐酸氢吗啡酮在膝关节镜手术术后镇痛中的临床应用

目的 评价关节腔内注射不同剂量盐酸氢吗啡酮对膝关节镜手术术后镇痛的效果. 方法 择期硬膜外麻醉下行膝关节镜手术患者150例,按随机数字表法分为5组(每组30例):H1组、H2组、H3组、M组、C组.术毕H1组关节腔内注射盐酸氢吗啡酮0.1 mg(用生理盐水配制成10ml),H2组关节腔内注射盐酸氢吗啡酮0.2 mg,H3组关节腔内注射盐酸氢吗啡酮0.3 mg,M组关节腔内注射吗啡2 mg,C组关节腔内注射等量生理盐水10ml.记录术后4、6、8、12、24 h患者在屈膝关节90°状态下的VAS,记录术后24h内需要追加镇痛药物的患者例数以及术后副作用的发生情况. 结果 术后8h内H1组、H2组、H3组、M组VAS评分比较,差异无统计学意义(P＞0.05);术后12 h VAS评分,H2组(2.5±0.6)分、H3组(2.1±0.7)分、M组(2.3±0.8)分低于H1组(3.1±0.6)分,差异有统计学意义(P＜0.01);术后24 h VAS评分,H3组(2.2±0.5)分低于H2组(3.1±0.8)分和M组(3.0±0.6)分,差异有统计学意义(JP＜0.05).术后需要追加镇痛药物的患者例数随盐酸氢吗啡酮剂量的增加而减少.结论 关节腔内注射不同剂量盐酸氢吗啡酮和吗啡用于膝关节镜均能获得良好的术后镇痛效果,0.3 mg盐酸氢吗啡酮的镇痛效果更佳.     

Involvement of hypothalamic histamine H1 receptor in the regulation of feeding rhythm and obesity

Histamine H(1) receptors (H(1)-Rs) are found in peripheral tissues and in regions of the hypothalamus that are concerned with regulating body composition. In the present study, we investigated the detailed mechanisms of histamine H(1)-Rs in the development of obesity. Histamine H(1)-R knockout (H1KO) mice gradually developed mature-onset obesity, which was accompanied by hyperphagia and decreased expression of uncoupling protein-1 (UCP-1) mRNA. Both younger nonobese (12-week-old) and older obese (48-week-old) H1KO mice exhibited impairment of the responsiveness to the leptin. In addition, disruption of the diurnal rhythm of feeding occurred before the onset of obesity in H1KO mice. Correction of these abnormal feeding rhythms by means of scheduled feeding caused a reduction in obesity and associated metabolic disorders in H1KO mice. Furthermore, central administration of a histamine H(1)-R agonist affected feeding behavior, body weight, and c-fos-like immunoreactivity in the hypothalamus. Taken together, these findings suggest that histamine H(1)-Rs are crucial for the regulation of feeding rhythm and in mediating the effects of leptin. Early disruption of H(1)-R-mediated functions in H1KO mice may lead to hyperphagia and decreased expression of UCP-1 mRNA, which may contribute to the development of obesity in these animals. In addition, centrally acting histamine H(1)-R may be a novel therapeutic target for the treatment of obesity and related metabolic disorders.     

PEP-1-血红素加氧酶-1融合蛋白转导对大鼠H9c2心肌细胞缺氧复氧损伤的影响

目的 探讨PEP-1-血红素加氧酶(HO)-1融合蛋白转导对大鼠H9c2心肌细胞缺氧复氧损伤的影响.方法 构建含人HO-1基因的原核表达质粒pETl5b-PEP1-hHO-1,质粒转化后诱导目的 蛋白PEP-1-HO-1表达.用含15%胎牛血清高糖DMEM培养基培养H9c2心肌细胞,随机分为4组(n=4):正常对照组(C组)常规培养;缺氧复氧组(H/R组)细胞缺氧22 h,复氧8 h;低浓度融合蛋白组(L-HO组)缺氧前用终浓度为1.0 μmol/L PEP-1-HO-1融合蛋白孵育细胞;高浓度融合蛋白组(H-HO组)缺氧前即刻用终浓度为2.0 μmol/L PEP-1-HO-1融合蛋白孵育细胞.复氧结束后收集细胞及培养液上清,采用2,4-二硝基苯肼显色法检测培养液乳酸脱氢酶(LDH)活性,硫代巴比妥酸比色法检测细胞MDA含量,黄嘌呤氧化酶法检测细胞SOD活性.结果 与C组比较,H/R组、L-HO组、H-HO组心肌细胞SOD活性降低,MDA含量升高,培养液LDH活性升高(P＜0.05);与H/R组比较,L-HO组和H-HO组心肌细胞SOD活性升高,MDA含量降低,培养液LDH活性降低(P＜0.05);与L-HO组比较,H-HO组心肌细胞SOD活性升高,MDA含量降低,培养液LDH活性降低(P＜0.05).结论 PEP-1-HO-1融合蛋白转导入大鼠H9c2心肌细胞可减轻细胞缺氧复氧损伤.