On the Impairment of Stress-Induced Changes in Triglyceride Levels via a Sub-Toxic Dose of Unmethylated Cytidine Phosphate Guanosine Oligodinucleotide (a Toll-Like Receptor 9 Ligand)

Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidine phosphate guanosine (CpG) oligodinucleotide (ODN1826), a Toll-like receptor 9 (TLR9) agonist was administered in combination with D-galactosamine (GalN) that caused relatively liver-specific inflammation and toxicity. In the control mice group injected with phosphate-buffered saline (PBS) (acute psychological stress model associated with blood sampling), the serum triglyceride (TG) levels showed a rapid decrease followed by a rebound at 24 h as we have recently reported. However, such a TG rebound was impaired in the CpG/GalN- and solely CpG-treated groups of mice despite an absence of liver injury based on serum alanine aminotransferase levels in the latter group. Thus, the stress-associated serum TG rebound was abrogated by the injection of a sub-hepatotoxic CpG dose. In the second experiment, we simply measured the hepatic CD36 and SACRB1 (the gene for scavenger receptor B1 (SR-B1)) transcripts after the i.p. administration of PBS, CpG or CpG/GalN. There was a remarkable elevation of hepatic CD36 transcript expression in both the CpG- and CpG/GalN-treated mice at 8 h post-CpG injection whereas the increase in the PBS-treated mice was slower than the former two groups, suggesting that hepatic CD36 transcript expression is more pronounced in the combined stress models than under psychological stress alone. The individual mice data showed that the increase in CD36 expression was accompanied by a reduction in SCARB1 mRNA, showing reciprocal regulation between these two genes. Together with our previously reported findings, these data suggest that, in a murine model combining psychological stress with TLR-triggered hepatic inflammation, the psychological stress facilitates liver uptake of plasma TG (and its components fatty acids), but the subsequent re-esterification and/or release of TG-rich lipoproteins from the liver is impaired due to the concomitant TLR-signaling. We hypothesize that lipid metabolism during acute stress shifts toward an elevated hepatic uptake of lipids due to concomitant TLR signaling, facilitating the clearance of bacterial lipids by the liver.

Changes in the gene expression of activation-induced cytidine deaminase in peripheral blood mononuclear cells in COVID-19 patients

Objective:To assess the changes in gene expression of activation-induced cytidine deaminase(AID)in COVID-19 patients and its relationship with the presence of autoantibodies in those ones.Methods:This retrospective study was carried out among COVID-19 patients’serum and peripheral blood mononuclear cells that were archived at the main COVID-19 referral centers in Iran from January to March 2022.The titer of autoantibodies in the serum of healthy and COVID-19 subjects was investigated by serological tests.Then,the AID gene expression in peripheral blood mononuclear cells in two groups was evaluated using the real-time polymerase chain reaction method.Results:100 patients were included.In total,8%and 1%of patients had a significant titer of anti-citrullinated protein antibody and antinuclear antibody autoantibodies,respectively,and those suffered from at least one comorbidity.No patient was positive for antiphospholipid autoantibody.Moreover,no significant changes were observed in the titer of autoantibodies between healthy subjects and patients.Furthermore,the increase in the expression of the AID gene was not significant in two groups.Conclusions:The presence of autoantibodies in COVID-19 patients might be linked to the previous underlying disease.

A spectral study of the interaction between chelerythrine chloride and cytidine

To study the possible anticancer mechanisms of chelerythrine (CHE), and its interactions with cytidine were investigated by UV–vis spectrophotometric and spectrofluorimetric measurements and by thermodynamic calculations. The binding of CHE to cytidine could be characterized by the hypochromic and bathochromic effects in the absorption bands, and the quenching of fluorescence intensity. The spectral data were fit by linear analysis, yielding a binding constant of 2.49 × 10(4) L mol(?1)at 25 °C of CHE and cytidine, and a van’t Hoff enthalpy of ?20.02 kJ/mol for the exothermic interaction in the standard state. In addition, with and , the interactions should be entropy-driven.

Symptomatic improvement in an acute, non-traumatic spine pain model with a combination of uridine triphosphate, cytidine monophosphate, and hydroxocobalamin

Rationale: In a previously published trial on spinal acute non-traumatic pain, peripheral neuro- regenerative combination of UTP, CMP and hydroxocobalamin presented unexpected analgesicproperties. Objective: To corroborate analgesiceffects of UTP, CMP and hydroxocobalamin combination in a self-paired evolutionary model. Methods: Mean VAS scores from pretreatment, V2 (5th treatment day) and V3 (10th treatment day) were plotted and statistically analyzed (ANOVA) for differences. PFQ scores from pretreatment, V2, and V3 were analyzed using the chisquare test. Results: The difference between V3 and pretreatment mean VAS scores was statistically significant (p < 0.0001). The improvement in PFQ scores throughout the study was found to be statistically significant (p < 0.0001). Conclusion: The combination of UTP, CMP and hydroxocobalamin seems to have analgesic properties in mediumterm use. The complex peripheral neu-roregenerative pharmacodynamics of this combination provides a plausible basis for this finding. Further randomized studies are needed to explore this combination for the indication of neuropathic pain due to spinal structure involvement.

Efficacy of Cytidine-5'-diphosp-bocholine Combined with Compound Anisodine in the Treatment of Early Optic Nerve Contusion

Purpose:To investigate the efficacy of anisodine combined with cytidine-5'-diphosp-bocholine (citicoline) in the treatment of early optic nerve contusion.Methods:A total of 33 subjects eligible for inclusion were selected from 105 patients clinically diagnosed with optic nerve contusion.These patients were subsequently divided into the control group (n =16) and the intervention group (n =17).In the control group,the participants received therapy consisting of glucocorticoids,mannitol,vasodilators and vitamin B.The patients in the intervention group additionally received anisodine in combination with citicoline.The visual acuity was graded on a scale from 0 to 8.Results:Prior to treatment,.the 25th,50th and 75th percentiles of visual acuity grade were 3,4 and 6.75 for the controls,and 3,4 and 6.5 for the patients in the intervention group (P=0.97).After treatment,the 25th,50th and 75th percentiles of visual acuity grade were 4,6 and 7.75 in the control group,and 7,7 and 8 in the intervention group (P=0.046).A significant difference was observed in both control (P=0.005) and intervention groups(P=0.001) when comparing presenting visual acuity before and after treatment.Conclusion:The combination of anisodine and citicoline with standard steroid and mannitol therapy appears to be effective in the treatment of early optic nerve contusion.

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Cytidine 5'-monophosphate(5'-CMP)is an essential nucleotide for additives.In this study,enhanced production of 5'-CMP was realized by the transformation of cytidine using co-immobilized di-enzymes,uridine-cytidine kinase(UCK)and acetate kinase(AcK).The immobilization yield of the enzyme had a clear correlation with the surface charges as zeta potential(ξ).Among them,ε-polylysinefunctionalized sepharose(SA-EPL,ξ=9.31 m V)showed high immobilization yield(78.8%),which was4.9-fold than that of nitrilotriacetic acid functionalized sepharose(SA-NTA,ξ=-12.6 m V).The residual activity of affinity co-immobilized enzyme(EPL-Ni/EPL@Ac K-UCK)was higher than 70.6%after recycled 10 times.Thus,this study provides an effective approach for the production of 5'-CMP with the advantages of low adenosine 5'-triphosphate(ATP)consumption,reduced side reactions,and improved reusability by co-immobilized UCK and Ac K on the functionalized Sepharose.

Protection of a cytidine deaminase gene gainst toxicity of high dose chemotherapy in mice

Objective： To explore the feasibility of transfecting cytidine deaminase （CD） gene into mouse bone marrow cells in order to observe the drug resistance of high dose Ara-C and improve the tolerance of myelosuppression following combination chemotherapy. Methods： Human cytidine deaminase gene was transfected into mice bone marrow cells by retroviral vector. Resistant colony-forming unit granulocyte-macrophage （CFU-GM） assay was performed after the transfected mice bone marrow cells treated by the Ara-C. DNA was extracted from mice bone marrow cells. The drug resistant gene in mice bone marrow cells after transfection was detected by PCR. Results： Bone marrow cells of the donor mice cultured with the retroviral producer cells showed the drug resistant colonies and resistance to Ara-C, so did accept mice transplanted with the CD gene （CFU-GM of donor mice was 52%, χ^2 = 124.62, P 〈 0.01; accept mice was 54%, χ^2 = 126.26, P 〈 0.01, both compared with the contrast group）. The animal survival rate was significantly higher in gene transfected group than that of the control （χ^2= 7.42, P 〈 0.01）. CD gene of transfected bone marrow cells was confirmed by PCR. Conclusion： CD gene can be transfected into bone marrow cells of mice efficiently and increase the drug resistance to Ara-C.

X-ray Examination of the Complex Adenosine, Guanosine and Cytidine with UO2^2＋ Ions

Interaction of dioxouranitm （VI） UO2^2＋ ion with Adenosine-5＇-triphosphate, Guanosine-5＇-triphosphate and 3ytidine-5＇-triphosphate were obtained a complexs of Adenosine, Guanosine and Cytidine with uranium UO2^2＋ ions and X-ray nethod to explore these complexes.

植酸-胞嘧啶对DL-蛋氨酸粉尘爆燃火焰抑制特性研究

饲料及其添加剂粉尘具有较高的燃烧热,在生产过程中存在一定的爆燃风险,威胁生命财产安全,预混抑制剂是目前应用最为广泛的一种抑爆手段,但传统抑制剂不可食用,无法加入至饲料类粉尘实现抑爆。因此,以饲料主要添加剂DL-蛋氨酸(DLM)粉尘为研究对象,采用自主合成的营养价值高、绿色可食用生物质基植酸-胞嘧啶(PA-CY),研究了PA-CY对DLM粉尘爆燃火焰传播特性的影响,通过高速摄影和可视化竖直管道记录爆燃火焰传播过程并计算火焰速度,采用热电偶监测火焰温度变化。结果表明:随PA-CY质量分数的升高DLM爆燃火焰亮度持续下降,严重破坏了火焰结构,加入20%PA-CY后火焰峰值速度、平均速度和峰值温度由27.66 m/s、14.39 m/s、1014℃分别下降至13.83 m/s、6.88 m/s、540℃,下降比重达50.0%、52.2%和46.7%,且PA-CY质量分数达30%后粉尘无法被点燃,说明PA-CY抑制效果显著。此结果可为饲料类粉尘爆燃的防治提供理论支持。

一锅法全细胞催化合成胞苷三磷酸的研究

分别在大肠杆菌BL21(DE3)中构建胞苷激酶(CDK)与乙酸激酶(ACK)的共表达及融合表达体系和胞苷酸激酶(CMK)与乙酸激酶(ACK)的融合表达体系,其中融合菌株SUMO-CDK-L2-ACK与CMK-L2-ACK构成的双融合催化体系具有较好的催化活性。搭建了双融合菌全细胞一锅法催化合成胞苷三磷酸(CTP)反应体系,并对其温度、pH、金属离子、乙酰磷酸投加量及菌粉投加量等进行优化。结果表明,在优化反应体系中,反应75 min时CTP产量达到最高为108 mmol/L,总收率达86.4%,实现了CTP的一锅法高效合成。

胞二磷胆碱联合替格瑞洛治疗急性脑梗死伴昏迷患者的疗效观察

目的研究胞二磷胆碱联合替格瑞洛治疗急性脑梗死伴昏迷的预后效果。方法80例急性脑梗死伴昏迷患者,依据随机法分为对照组及观察组,各40例。对照组患者给予替格瑞洛治疗,观察组在对照组基础上联合胞二磷胆碱治疗。比较两组患者治疗效果、不良反应发生情况、昏迷评分、神经功能缺损评分、白细胞介素-6(IL-6)水平、白细胞介素-8(IL-8)水平。结果治疗后,观察组总有效率90.00%高于对照组的70.00%(P<0.05);两组不良反应发生率比较无统计学意义(P>0.05)。治疗后,两组患者的昏迷评分高于治疗前、神经功能缺损程度评分以及IL-6、IL-8水平均低于治疗前,观察组患者的昏迷评分(14.80±1.50)分高于对照组的(12.05±1.30)分,神经功能缺损程度评分(9.60±1.20)分以及IL-6(9.10±0.55)ng/ml、IL-8(2.85±0.20)ng/ml均低于对照组的(13.20±1.50)分、(12.30±1.20)ng/ml、(4.45±0.30)ng/ml(P<0.05)。结论胞二磷胆碱与替格瑞洛联合应用对急性脑梗死伴昏迷患者有确切的临床疗效,能够显著改善患者昏迷状态,修复损伤的神经功能,同时有较高的安全性。

左卡尼汀联合胞二磷胆碱对新生儿缺氧缺血性脑病的保护作用

目的 观察左卡尼汀(LC)联合胞二磷胆碱(CDPC)对新生儿缺氧缺血性脑病(HIE)的保护作用。方法 选取2019年8月至2023年8月青海省西宁市第二人民医院就诊的HIE患儿98例,根据随机数字表法将其分为对照组(CDPC治疗)、试验组(LC联合CDPC治疗),各49例。两组均治疗14 d,比较两组恢复情况、治疗效果及不良反应;比较两组治疗前后血清肌酸激酶同工酶-MB(CK-MB)、神经元特异性烯醇化酶(NSE)、S100B、内皮素-1(ET-1)、脂氧素A4(LXA4)水平。结果 试验组惊厥停止、反射恢复、肌张力恢复、意识恢复时间短于对照组(P<0.05)。试验组临床疗效优于对照组(P<0.05)。治疗后,两组血清CK-MB、NSE、S100B、ET-1水平低于治疗前,且试验组低于对照组(P<0.05);两组LXA4水平高于治疗前,且试验组高于对照组(P<0.05)。两组不良反应总发生率比较,差异无统计学意义(P>0.05)。结论 LC联合CDPC可加快HIE恢复,提升治疗效果,降低血清CK-MB、NSE、S100B、ET-1水平,提高LXA4水平,且安全可靠。

注射用三磷酸胞苷二钠细菌内毒素检查法的研究

目的建立注射用三磷酸胞苷二钠细菌内毒素检查方法。方法按《中国药典》2020年版四部通则1143细菌内毒素检查法,从严确定注射用三磷酸胞苷二钠的限值,用凝胶法进行方法学验证和细菌内毒素检查。结果两个厂家的4批供试品在0.2 mg/ml浓度下对两个厂家的鲎试剂不存在干扰,细菌内毒素的限值设定为2.5 EU/mg,4批注射用三磷酸胞苷二钠细菌内毒素的检查结果均符合规定。结论本研究建立的细菌内毒素检查方法可用于替代家兔热原检查法,满足对注射用三磷酸胞苷二钠的质量控制要求。

基于cdd基因敲除和嘧啶操纵子转移的胞苷产生菌的研究

以大肠杆菌作为出发菌株,通过基因工程手段对其进行改造,旨在提高胞苷产量.首先,通过Red重组系统敲除了大肠杆菌MG3028基因组上的胞苷脱氨酶基因(cdd),阻断了胞苷的分解代谢.然后,构建了含解淀粉芽孢杆菌TS8嘧啶操纵子结构基因的重组质粒pPYR3021,并将其转入E.coli MG3028(△cdd).与出发菌株E.coli MG3028相比,E.coli MG3028(△cdd)的胞苷产量提高了近2倍,而尿苷产量降低了近75%,携带重组质粒pPYR3021的菌株胞苷产量较出发菌株提高了近6倍.

结直肠癌术后辅助卡培他滨为基础化疗方案的临床疗效及相关药物基因组学分析

目的:本研究旨在探讨结直肠癌(colorectal cancer,CRC)术后辅助卡培他滨为基础化疗方案的疗效及相关药物基因组学分析。方法:回顾性分析2010年1月至2017年6月河南中医药大学第三附属医院收治术后接受辅助化疗的CRC患者215例。收集外周血提取DNA用来进行胞苷脱氨酶(cytidine deaminase,CDD)多态性位点基因分型。另外,收集85例患者的化疗前外周血单核细胞(peripheral blood mononuclear cell,PBMC)提取RNA对CDD的m RNA表达水平进行测定。结果:215例患者具有良好的疗效。药物基因组学分析方面,发现CDD基因启动子区域的多态性位点-451C>A和疗效相关,该位点GA/AA和GG基因型患者的3年无疾病生存率(disease-free survival,DFS)分别为81.03%和62.42%,差异具有统计学意义(P=0.002)。两种基因型患者的5年总生存率(overall survival,OS)分别为72.41%和53.50%,差异具有统计学意义(P=0.016)。在85例患者外周血单核细胞的mRNA表达分析中发现,携带A等位基因的患者相对于GG型患者,外周血单核细胞中CDD的mRNA表达明显较高,差异具有统计学意义(P<0.001)。结论:卡培他滨为基础的化疗方案具有良好的治疗效果。CDD基因-451C>A位点可能通过影响该基因m RNA的表达使CRC患者从卡培他滨的治疗当中获益。

三磷酸胞苷加地塞米松等综合治疗突发性聋的临床观察

目的 对三磷酸胞苷 (cyntridinitriphosphatis ,CTP)加地塞米松等综合治疗突发性聋的疗效进行临床观察。方法 将 2 0 7例 (312耳 )突发性聋患者按照就诊顺序分成CTP组 10 6例 (15 9耳 )和对照组 10 1例 (15 3耳 )。CTP组使用CTP治疗 ,对照组采用传统的ATP、辅酶A、肌苷 0 4g和维生素C治疗 ;两组还同时加用地塞米松、维生素B、吸氧、丹参等完全相同的治疗。如有明确的感冒受凉史(CTP组 2 5例 ,对照组 2 7例 ) ,则加用抗病毒药及干扰素。两组在治疗前和治疗后 2d开始每 4d用纯音电测听测试患者的听力。结果 CTP组和对照组在治疗前平均听阈分别为 (76 5 6± 19 0 4 )dBHL和 (74 0 1± 17 0 7)dBHL(P >0 0 5 ) ;治疗后 ,CTP组 10 6例中痊愈 6 7例 (94耳 )、显效 2 1例 (35耳 )、有效 16例 (2 7耳 )及无效 2例 (3耳 ) ,治愈率 5 9 12 % ,平均听阈为 (43 0 8± 2 1 4 7)dBHL ,听力平均提高(2 7 39± 18 4 4 )dB ,未见任何副作用 ;对照组 10 1例 (15 3耳 )中痊愈 16例 (2 5耳 )、显效 19例 (32耳 )、有效 5 1例 (73耳 )及无效 15例 (2 3耳 ) ,治愈率 16 34% ,平均听阈分别为 (5 7 13± 32 4 1)dBHL ,平均听力提高 (19 4 5± 19 12 )dB。两组经统计学处理差异有显著性 (P <0 0 5 )。

枯草芽孢杆菌cdd基因敲除及对胞苷发酵的影响

目的:通过敲除出发菌株上的胞苷脱氨酶基因,阻断嘧啶代谢通量由胞苷流向尿苷和尿嘧啶,选育胞苷产生菌。方法:采用同源重组的方法敲除枯草芽孢杆菌TS8的胞苷脱氨酶基因cdd,并通过遗传稳定性实验验证其缺失标记和胞苷产量,通过摇瓶发酵实验对比出发菌株和缺失株的产苷水平。结果:cdd基因缺失菌株TSb发酵72h,发酵液中胞苷产量达到1.72g/L,与原始菌株相比提高了44.19%,且遗传性状稳定。结论:cdd基因的缺失可有效阻断嘧啶代谢通量由胞苷流向尿苷和尿嘧啶,提高胞苷产量。

大肠杆菌cdd和thrA基因的敲除及其对胞苷积累量的影响

以大肠杆菌AU39作为出发菌株,通过基因工程手段对其进行改造,旨在提高胞苷产量。首先,通过Red重组系统敲除了大肠杆菌AU39基因组上的胞苷脱氨酶基因cdd,阻断了胞苷的分解代谢;敲除了E.coli AU39(Δcdd)基因组上的高丝氨酸脱氢酶基因thrA,阻断天冬氨酸向高丝氨酸的代谢途径。然后,分别对不同基因缺失菌株进行培养发酵,与出发菌株E.coli AU39相比,两株突变株都有不同程度的胞苷积累,E.coli AU39(Δcdd)与E.coli AU39(ΔcddΔthrA)的胞苷产量提高了1.25倍与1.6倍,而尿苷产量均相对有所降低。

利用啤酒酵母生物合成胞苷三磷酸的研究

研究了利用游离的啤酒酵母将胞苷一磷酸(CMP)合成为胞苷三磷酸(CTP),采用HPLC法分析反应过程中各工艺参数对转化率的影响并对其主要因素进行了正交优化。结果表明,在25g酵母量中加入20mL底物溶液,其中包括CMP60mmol/L、葡萄糖150mmol/L、磷酸盐缓冲液(钠盐)250mmol/L、氯化镁8mmol/L、pH为6.0,温度35℃、反应时间2.0h,在此反应体系下,通过添加适量ATP、吐温,CTP的转化率可维持在80%以上。

HPLC-MS^2鉴定大青叶水提液中的5种化学成分

目的:鉴定大青叶水提液中的化学成分。方法:大青叶的水提液经HPLC分离后,收集各色谱峰的馏分进行二级质谱分析,并和胞苷、尿苷、鸟苷、黄嘌呤和次黄嘌呤对照品的水溶液进行比较。结果:大青叶水提液HPLC中的5个色谱峰分别与胞苷、尿苷、鸟苷、黄嘌呤和次黄嘌呤对照品的t_R和UV光谱一致,并具有相同的二级质谱碎片离子。结论:大青叶中含有胞苷、次黄嘌呤、尿苷、黄嘌呤和鸟苷等5种成分。