不同诱导子对枇杷悬浮细胞生长及熊果酸、齐墩果酸含量的影响

在已建立的稳定枇杷细胞悬浮培养系的试验中,研究加入诱导子水杨酸（SA）、茉莉酸甲酯（MJ）、水解酪蛋白（CH）和酵母提取物（YE）对枇杷悬浮细胞生长及次生代谢产物熊果酸（UA）、齐墩果酸（OA）含量的影响。结果表明,从细胞生长、UA及OA含量三方面综合考虑,以MJ为100.0 mg.L^-1时培养最佳,此时细胞生长、UA及OA含量分别为对照的1.081、2.540及2.590倍。     

Hernandulcin Production in Cell Suspensions of Phyla Scaberrima: Exploring Hernandulcin Accumulation through Physical and Chemical Stimuli

Hernandulcin (HE) is a non-caloric sweetener synthesized by the Mexican medicinal plant Phyla scaberrima. Herein we present the results of HE production through cell suspensions of P. scaberrima as well as the influence of pH, temperature, biosynthetic precursors and potential elicitors to enhance HE accumulation. The incorporation of mevalonolactone (30–400 mg L⁻¹) farnesol (30–400 mg L⁻¹), AgNO3 (0.025–0.175 M), cellulase (5–60 mg L⁻¹; 0.3 units/mg), chitin (20–140 mg L⁻¹) and (+)-epi-α-bisabolol (300-210 mg L⁻¹) to the cell suspensions, resulted in a differential accumulation of HE and biomass. Among elicitors assayed, chitin, cellulase and farnesol increased HE production from 93.2 to ∼160 mg L⁻¹ but, (+)-epi-α-bisabolol (obtained by a synthetic biology approach) increased HE accumulation up to 182.7 mg L⁻¹. HE produced by the cell suspensions was evaluated against nine strains from six species of gastrointestinal bacteria revealing moderate antibacterial activity (MIC, 214–465 μg mL⁻¹) against Staphylococcus aureus, Escherichia coli and Helicobacter pylori. Similarly, HE showed weak toxicity against Lactobacillus sp. and Bifidobacterium bifidum (>1 mg mL⁻¹), suggesting a selective antimicrobial activity on some species of gut microbiota. According to our results, chitin and (+)-epi-α-bisabolol were the most effective molecules to enhance HE accumulation in cell suspensions of P. scaberrima.     

前体、诱导子及抑制剂对紫杉烷生物合成的促进作用研究

研究了前体、诱导子和抑制剂对中国红豆杉细胞培养生产紫杉烷的促进作用。结果表明,向培养基中加入30mg/L 3-甲基－2－丁烯－1－醇,2mmol/L苯甲酸钠,10mg/L矮壮素,100μmol/L茉莉酸甲酯,0.1mmol/L丝氨酸,可以使紫杉醇含量增加1141.1%;加入0.1mmol/L苯甲酸钠,80μmol/L茉莉酸甲酯,0.1mmol/L丝氨酸可以使2α,5α,10β,14β－四乙酰氧基－紫杉－4（20）,11－二烯含量增加134.6%;25mg/L矮壮素,100μmol/L茉莉酸甲酯,0.5mmol/L丝氨酸可以使1β－羟基巴卡亭I含量增加95.2%;5mg/L3－甲基－2－丁烯－1－醇,10mg/L矮壮纱,40μmol/L茉莉酸甲酯,可以使14β-(2-甲基丁酰氧基）－2α,5α,10β－三乙酰氧基－紫杉－4（20）,11－二烯的含量增加76.4%。     

Resistance to Pseudorabies Virus with Enhanced Interferon Production and Natural Killer Cell Activity in Mice Treated with Serum Thymic Factor

Susceptibility to pseudorabies virus (PRV) infection in mice, which were continuously depleted of natural killer (NK) cell activity by injection of anti-asialo GM1, was examined. Effects of serum thymic factor (FTS) on susceptibility of mice to PRV were also investigated. In mice with depleted NK cell activity, the mortality of PRV-infected mice was markedly high, whereas that of FTS-pretreated mice was significantly lower than the controls. Reduced susceptibility to PRV was demonstrated in mice treated with anti-asialo GM1 antisera before the PRV infection. Such a reduced susceptibility was not observed in mice inoculated with the antisera on day 1 post-infection (PI). To analyze the FTS-induced resistance to PRV infection, NK cell activity, macrophage activity, and interferon (IFN) productions were studied. Interferon production and NK cell activity were enhanced in the FTS-pretreated mice, suggesting that interferon may play an important role in this FTS-induced resistance to PRV infection.     

两株毒力不同的核盘菌产草酸、果胶酶的比较

对强弱毒力不同的核盘菌Ep-1PNA5和Ep-1PN的主要致病因子草酸和果胶酶的产生进行了比较研究。结果发现强毒力的Ep-1PNA5和弱毒力Ep-1PN都可产生草酸,并且在发病油菜活体组织上,弱毒力Ep-1PN的病组织中的草酸含量高于Ep-1PNA5病组织;两个菌株在发病油菜活体组织上的果胶酶产量没有差异,但在诱导培养基中Ep-1PN菌株比Ep-1PNA5的果胶酶产量高。这一结果表明弱毒力Ep-1PN菌株毒力的衰退并不是因为其所携带的dsRNA因子抑制了草酸和果胶酶所产生的。     

Salicylic Acid Enhances Jaceosidin and Syringin Production in Cell Cultures of Saussurea medusa

Addition of 20 μM salicylic acid to Saussurea medusa cell cultures at day 6 resulted in jaceosidin and syringin productions up to 95 mg l⁻¹ and 631 mg l⁻¹ which were, respectively, about 2.5- and 2.7-fold higher than in the control. The biomass was increased from 8 to 12 g l⁻¹. Expression of chalcone synthase gene (chs) increased sharply after 12 h treatment and was sustained up to 48 h; chalcone isomerase gene (chi) expression reached a peak at 24 h and decreased after 48 h; and phenylalanine ammonia-lyase activity increased by 7.5-fold (96 U mg⁻¹ protein) higher than in the control after 24 h. These results indicate that salicylic acid enhances the production of jaceosidin and syringin which is accompanied by induction of the related phenylpropanoid biosynthetic enzymes.     

Effects of Fusaric Acid on Reactive Oxygen Species and Antioxidants in Tomato Cell Cultures

Generation of O₂^– and H₂O₂ as well as the activities of superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase, dehydroascorbate reductase and ascorbate content were studied in tomato cell cultures in response to fusaric acid – a nonspecific toxin of phytopathogenic Fusarium species. Toxin treatment resulted in decreased cell viability which was preceded by culture medium alkalinization up to 0.65 pH unit and enhanced extracellular O₂^– production. The H₂O₂ level was not significantly affected. In toxin-treated cultures, a transient, significant increase occurred in intracellular superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase activities. Fusaric acid-induced ascorbate turnover modulation led to up to a twofold increase in dehydroascorbic acid accumulation, and a decrease in the associated ascorbate redox ratio. It was concomitant with a significant decrease in dehydroascorbate reductase activity. These results support previous observations that the pro- and anti-oxidant systems are involved in response to fusaric acid treatment although differential response of H₂O₂ and its metabolism-related enzymes between the whole leaf and cell culture assays was found.     

Epilepsy, Neurodegeneration, and Extracellular Glutamate in the Hippocampus of Awake and Anesthetized Rats Treated with Okadaic Acid

We have previously shown that the intrahippocampal microinjection of okadaic acid (OKA), a potent inhibitor of serine/threonine protein phosphatases, induces epileptic seizures, neuronal death, and the hyperphosphorylation of the NR2B subunit of the N-methyl-d-aspartate (NMDA) receptor. We administered OKA by reverse microdialysis in the hippocampus of awake and halothane-anesthetized rats, with simultaneous collection of microdialysis fractions and recording of the EEG activity, and subsequent histological analysis. OKA produced intense behavioral and persistent EEG seizure activity in the awake rats but not in the anesthetized animals, and did not significantly alter the extracellular concentration of glutamate and aspartate detected in the microdialysis fractions. One day after the experiment a remarkable neurodegeneration of CA1 hippocampal region was observed in both the awake and the anesthetized rats. We conclude that the OKA-induced epilepsy cannot be ascribed to increased extracellular glutamate, but to an increased sensitivity of NMDA receptor. We propose that halothane protected against the epilepsy because it blocks NMDA receptor overactivation, and that the neurodegeneration of CA1 region is independent of this overactivation and due probably to alterations of cytoskeletal proteins consequent to the OKA-induced hyperphosphorylation.     

磷脂酶D对ParA1诱导的烟草的细胞死亡和过氧化氢及莨菪亭积累的影响

利用药理学方法,研究了烟草寄生疫霉(Phytophthora parasitica)分泌的蛋白激发子ParA1诱导烟草悬浮细胞后,磷脂酶D对ParA1诱导的过敏细胞死亡和其它防卫反应的影响.用100nmol/LParA1处理烟草悬浮细胞后能够诱导细胞死亡、过氧化氢和莨菪亭的积累.磷脂酶D抑制剂正丁醇能够抑制ParA1诱导的这些防卫反应,仲丁醇所起的抑制作用比正丁醇小,正丁醇和仲丁醇产生的抑制效果具有浓度依赖效应.而叔丁醇不能抑制ParA1诱导的这些反应.结果表明,磷脂酶D参与了ParA1诱导烟草悬浮细胞的信号传导过程.     

Purification,Cloning and Functional Expression of hydroxyphenylpyruvate Reductase Involved in Rosmarinic Acid Biosynthesis in Cell Cultures of Coleus Blumei

Hydroxyphenylpyruvate reductase (HPPR) is an enzyme involved in the biosynthesis of rosmarinic acid in Lamiaceae reducing hydroxyphenylpyruvates in dependence of NAD(P)H to the corresponding hydroxyphenyllactates. The HPPR protein was purified from suspension cells of Coleus blumei accumulating high levels of rosmarinic acid by ammonium sulfate precipitation, anion exchange chromatography, hydroxylapatite chromatography, chromatography on 2',5'-ADP-Sepharose 4B and SDS-polyacrylamide gel electrophoresis. The protein was tryptically digested and the peptides sequenced. Sequence information was used to isolate a full-length cDNA-clone for HPPR (EMBL accession number AJ507733) by RT-PCR, screening of a C. blumei cDNA-library and 5'-RACE-PCR. The open reading frame of the HPPR-cDNA consists of 939 nucleotides encoding a protein of 313 amino acid residues. The sequence showed that HPPR belongs to the family of D-isomer-specific 2-hydroxyacid dehydrogenases. The HPPR-cDNA was heterologously expressed in Escherichia coli and the protein was shown to catalyse the NAD(P)H-dependent reduction of 4-hydroxyphenylpyruvate to 4-hydroxyphenyllactate and 3,4-dihydroxyphenylpyruvate to 3,4-dihydroxyphenyllactate.     

咖啡酸对食管鳞状细胞癌KYSE450裸鼠移植瘤生长的影响及分子机制研究

摘要 目的：探讨与研究咖啡酸对食管鳞状细胞癌KYSE450裸鼠移植瘤生长的影响及分子机制。方法：将食管鳞状细胞癌移植瘤裸鼠（n=48）随机平分为三组-模型组、5-氟尿嘧啶组与咖啡酸组。三组分别经腹腔注射0.2 mL生理盐水、5-氟尿嘧啶25 g/kg、5-氟尿嘧啶25 g/kg与咖啡酸50 mg/kg,2次/周,持续4周。结果：5-氟尿嘧啶组与咖啡酸组治疗第2周与第4周的体重高于模型组（P<0.05）,咖啡酸组高于5-氟尿嘧啶组（P<0.05）。5-氟尿嘧啶组与咖啡酸组治疗第2周与第4周的肿瘤体积少于模型组（P<0.05）,咖啡酸组高于5-氟尿嘧啶组（P<0.05）。5-氟尿嘧啶组与咖啡酸组治疗第4周的血清TNF-α与IL-6含量低于模型组（P<0.05）,咖啡酸组低于5-氟尿嘧啶组（P<0.05）。5-氟尿嘧啶组与咖啡酸组治疗第4周的移植瘤Bax、Caspase-3蛋白相对表达水平与凋亡指数高于模型组（P<0.05）,咖啡酸组高于5-氟尿嘧啶组（P<0.05）。结论：咖啡酸在食管鳞状细胞癌裸鼠的应用能与5-氟尿嘧啶发挥协同作用,能通过上调Bax、Caspase-3蛋白的表达,促进移植瘤细胞凋亡,抑制炎症因子的表达,减少血管总数,从而抑制移植瘤生长,促进恢复裸鼠体重。     

肺功能康复训练联合肠外氨基酸营养支持对非小细胞肺癌化疗患者营养状态、癌因性疲乏及生活质量的影响

摘要 目的：探讨肺功能康复训练联合肠外氨基酸营养支持对非小细胞肺癌（NSCLC）化疗患者营养状态、癌因性疲乏及生活质量的影响。方法：选取2016年6月~2019年10月期间本院收治的NSCLC化疗患者137例。根据随机数字表法将患者分为对照组（68例,给予常规干预）、观察组（69例,在对照组基础上给予肠外氨基酸营养支持联合肺功能康复训练）,对比两组干预前后的营养状态、肺功能、癌因性疲乏及生活质量。结果：与对照组相比,观察组干预后白蛋白、总蛋白、血红蛋白更高（P<0.05）。干预后观察组用力肺活量（FVC）、第1s用力呼气容积占预计值百分比（FEV₁%）、呼气峰流速值（PEF）高于对照组（P<0.05）。观察组干预后Piper 疲乏量表（PFS）的行为疲乏、情感疲乏、躯体疲乏、认知疲乏评分低于对照组（P<0.05）。观察组干预后生活质量评估量表（EORTC QLQ-C30）各维度评分较对照组更高（P<0.05）。结论：肺功能康复训练联合肠外氨基酸营养支持干预NSCLC化疗患者,可改善患者的营养状态及肺功能,缓解患者癌因性疲乏程度,改善其生活质量。     

不同化疗方案对晚期非小细胞肺癌患者骨髓抑制及免疫功能的影响

目的:探讨不同化疗方案对晚期非小细胞肺癌患者骨髓抑制及免疫功能的影响。方法:选取2011年1月至2013年12月收治的130例晚期非小细胞肺癌患者,按照知情同意原则随机分为三组:NP(长春瑞滨+顺铂)组45例、GP(吉西他滨+顺铂)组43例、TP(紫杉醇+顺铂)组42例,分别于化疗前及化疗2个周期后检测患者的骨髓抑制及免疫水平。结果:三种化疗方案进行治疗后骨髓抑制水平由高到低排列为GP组、TP组、NP组,差异有统计学意义(P0.05);GP组血小板减少发生率高于其他两组,TP组白细胞下降发生率高于其他两组,差异有统计学意义(P0.05);三组患者化疗后的免疫功能指标均较化疗前低,差异有统计学意义(P0.05)。三种化疗方案进行治疗后免疫功能抑制水平由高到低排列为GP组、TP组、NP组,差异有统计学意义(P0.05)。结论:GP组患者血小板下降更明显,TP组白细胞下降更为明显,NP组对骨髓抑制及免疫功能抑制较缓和,更适于老年人,因此临床选择化疗方案时要综合考虑患者骨髓状况、免疫功能情况及年龄等。     

甘油三酯葡萄糖乘积指数、红细胞分布宽度、血清尿酸、降钙素原与冠心病患者冠状动脉狭窄程度和短期预后的关系分析

摘要 目的：探讨甘油三酯葡萄糖乘积（TyG）指数、红细胞分布宽度（RDW）、血清尿酸（UA）、降钙素原（PCT）与冠心病患者冠状动脉狭窄程度和短期预后的关系。方法：选取2017年1月至2021年12月我院收治的150例冠心病患者,均行冠状动脉造影并以Gensini积分系统评估冠状动脉狭窄程度,分为轻度组、中度组和重度组,比较各组的TyG指数、RDW、UA、PCT水平,以Pearson相关系数分析上述指标与Gensini积分的相关性。患者出院后均随访6个月,根据有无发生主要心血管不良事件（MACE）分为预后良好组和预后不良组,以多因素Logisitc回归分析预后的影响因素,以受试者工作特征（ROC）曲线分析对预后的预测效能。结果：不同冠状动脉狭窄程度患者的TyG指数、RDW、UA、PCT水平差异均表现为,从轻度到重度依次升高（P＜0.05）。Pearson相关性分析显示,冠心病患者的TyG指数、RDW、UA、PCT水平与Gensini积分均呈正相关（P＜0.05）。预后不良组的甘油三酯（TG）、Gensini积分、TyG指数、RDW、UA、PCT水平均高于预后良好组（P＜0.05）;多因素Logistic回归分析显示,TG≥3.5 mmol/L、Gensini积分≥30分、TyG指数≥5、RDW≥12.8%、UA≥380 μmol/L、PCT≥35 μg/L是冠心病患者预后不良的危险因素（P＜0.05）。ROC曲线分析显示,TyG指数、RDW、UA、PCT单独及联合预测冠心病患者预后不良的曲线下面积分别为0.786、0.793、0.794、0.789和0.948,联合预测效能明显更高。结论：冠心病患者的TyG指数、RDW、血UA、PCT水平与冠状动脉狭窄程度呈正相关,其均为患者短期预后的影响因素,联合检测上述指标对患者短期预后有一定预测价值。     

不同化疗方案联合手术治疗非小细胞肺癌的临床疗效及对患者免疫功能的影响分析

目的:探讨分析不同化疗方案联合手术治疗非小细胞肺癌的临床疗效以及对患者免疫功能的影响。方法:经病理学证实的非小细胞肺癌患者94例,随机数字表法将其分为顺铂组与洛铂组,各47例。2组患者均采取手术治疗,并术前行辅助化疗。其中顺铂组采取紫杉醇+顺铂方案,洛铂组接受紫杉醇+洛铂方案。比较治疗前后2组患者临床疗效及KPS评分,并对Ig A、Ig G、Ig M、CD4+及CD8+指标水平进行分析。结果:洛铂组有效率(CR+PR)为87.2%(41/47),与顺铂组[80.8%(38/47)]差异无统计学意义(P0.05)。洛铂组术后KPS评分及CD4+水平高于顺铂组(P0.05),但CD8+水平低于顺铂组(P0.05),差异有统计学意义;2组患者手术前后Ig A、Ig G及Ig M均无统计学意义(P0.05)。洛铂组毒副反应发生率低于顺铂组(P0.05)。结论:含洛铂术前辅助化疗方案有助于改善非小细胞肺癌患者的免疫功能,降低化疗的毒副反应。     

高尿酸血症患者血清尿酸水平与糖耐量异常、炎症因子及红细胞参数的相关性分析

摘要 目的：探讨高尿酸血症（HUA）患者血清尿酸（UA）水平与糖耐量异常、炎症因子及红细胞参数的相关性。方法：选取2020年1月~2021年1月期间我中心在健康体检中诊断的HUA患者240例（观察组）,另选取同期在我中心体检的健康志愿者200例（对照组）,分别检测血清UA、炎症因子：超敏C反应蛋白（hs-CRP）、单核细胞趋化蛋白1（MCP-1）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、红细胞参数：红细胞分布宽度（RDW）、红细胞平均体积（MCV）及红细胞计数、糖耐量指标：空腹血糖（FPG）、餐后2h血糖（2hPG）及空腹胰岛素（FINS）,计算稳态模型胰岛素抵抗指数（HOMA-IR）和稳态模型胰岛素敏感指数（HOMA-ISI）。结果：观察组的UA、RDW及红细胞计数均高于对照组,MCV低于对照组（P<0.05）。观察组的hs-CRP、MCP-1、IL-6及TNF-α均高于对照组（P<0.05）。观察组的FPG、2hPG、FINS及HOMA-IR均高于对照组,HOMA-ISI低于对照组（P<0.05）。Pearson相关系数指出,HUA患者血清UA水平与红细胞计数、RDW、hs-CRP、MCP-1、IL-6、TNF-α、FPG、2hPG、FINS及HOMA-IR呈正相关,而与HOMA-ISI、MCV呈负相关（P<0.05）。结论：高尿酸水平可引起HUA患者炎症反应增强、糖耐量异常及红细胞参数改变,控制UA水平可减少多种并发症的发生风险。     

不同类型冠心病患者血清Hcy、TBIL、hs-CRP、尿酸的表达及临床意义

目的:探讨不同类型冠心病患者血清同型半胱氨酸(Hcy)、总胆红素(TBIL)、高敏C反应蛋白(hs-CRP)、尿酸的水平及临床意义。方法:选取首都医科大学附属北京地坛医院2015年9月-2018年7月收治的冠心病患者132例为冠心病组,根据临床诊断分为稳定型心绞痛52例(SAP组)、不稳定型心绞痛42例(UAP组)、急性心肌梗死38例(AMI组),另选取50例同时期于我院体检的健康志愿者为对照组,检测各组Hcy、TBIL、hs-CRP、尿酸的水平,采用Pearson相关分析Hcy、TBIL、hs-CRP、尿酸水平之间的相关性,采用Logistic回归分析冠心病的影响因素。结果:冠心病组患者的血清Hcy、hs-CRP、尿酸水平显著高于对照组,TBIL水平显著低于对照组(P0.05)。AMI、UAP组患者的血清Hcy、hs-CRP、尿酸水平显著高于SAP组,TBIL水平显著低于SAP组(P0.05),且AMI组患者的血清Hcy、hs-CRP、尿酸水平显著高于UAP组,TBIL水平显著低于UAP组(P0.05)。经Pearson相关分析显示,Hcy与hs-CRP、尿酸呈正相关,与TBIL呈负相关,hs-CRP与尿酸呈正相关(P0.05),TBIL与hs-CRP、尿酸无明显相关性(P0.05)。经Logistic回归分析显示,Hcy、hs-CRP、尿酸、高血压、糖尿病均是冠心病的独立危险因素(P0.05),TBIL是冠心病的保护因素(P0.05)。结论:冠心病患者血清Hcy、hs-CRP、尿酸水平升高,TBIL水平降低,Hcy、TBIL、hs-CRP、尿酸与患者的病情相关,也是冠心病的影响因素。     

右美托咪定联合不同剂量咪达唑仑对非小细胞肺癌手术患者血流动力学和炎症介质、认知功能的影响

摘要 目的：探讨右美托咪定联合不同剂量咪达唑仑对非小细胞肺癌（NSCLC）手术患者血流动力学和术后炎症介质、认知功能的影响。方法：纳入2019年1月-2021年2月期间来郑州大学第一附属医院接受手术治疗的NSCLC患者（n=120）,按照麻醉方案的不同分为低剂量组（n=60）和高剂量组（n=60）,两组均予以右美托咪定,在此基础上,低剂量组予以0.05 mg/kg咪达唑仑,高剂量组予以0.10 mg/kg咪达唑仑。对比两组患者的血流动力学、炎症介质、认知功能、苏醒质量和不良反应发生率。结果：两组麻醉诱导后10 min（T1）~拔管后5 min（T3）时间点心率（HR）、平均动脉压（MAP）升高后下降,且拔管即刻（T2）、T3时间点高剂量组低于低剂量组（P<0.05）。高剂量组的苏醒后疼痛视觉模拟评分（VAS）评分低于低剂量组,睁眼时间、拔管时间、麻醉恢复室（PACU）停留时间均短于低剂量组（P<0.05）。两组术前、术后3 d、术后7 d白介素-1（IL-1）、C反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）升高后下降,且术后3 d、术后7 d高剂量组低于低剂量组（P<0.05）。两组术后1 d、术后3 d、术后7 d简易精神状态量表（MMSE）评分组间对比未见统计学差异（P>0.05）。两组不良反应发生率组间对比无统计学差异（P>0.05）。结论：与0.05 mg/kg咪达唑仑相比,右美托咪联合0.10 mg/kg咪达唑仑定用于NSCLC手术患者,维持血流动力学稳定、提高苏醒质量、减轻炎症应激的效果更为显著,且不增加认知功能损害和不良反应发生率。