慢性乙型肝炎及乙型肝炎性肝硬化患者色氨酸代谢变化及其相关性分析

目的 探讨慢性乙型肝炎(CHB)和乙型肝炎性肝硬化(HBC)患者色氨酸(Trp)代谢变化及相关性.方法 选取2015年10月至2016年1月在重庆医科大学附属第一医院住院的CHB 81例和HBC 40例患者为研究对象,40例健康者为对照.采用高效液相色谱-荧光检测法(HPLC-FD)对各组受试者血浆中Trp及代谢产物犬尿氨酸(Kyn)、5-羟色胺(5-HT)水平进行测定,实时荧光定量PCR检测HBV-DNA病毒复制数,分析3组间血清学指标差异,采用Pearson相关分析和Logistic回归分析筛选危险因素.结果 与对照组比较,CHB组、HBC组受试者血浆中Trp和5-HT水平明显降低(P<0.01);HBC组受试者血浆中Kyn/Trp明显高于其他两组(P<0.01).HBC组Kyn/Trp与A/G呈负相关(r=-0.686,P<0.01),CHB组Kyn/Trp与Log DNA呈正相关(r=0.784,P<0.01).Logistic回归分析显示Kyn/Trp为CHB和HBC的危险因素.结论 检测血浆中Trp及其代谢产物可作为辅助指标评估乙型肝炎进展情况.     

慢性乙型肝炎患者色氨酸和犬尿氨酸的检测及意义

目的探讨血浆中色氨酸(Trp)和犬尿氨酸(Kyn)的定量检测在乙型肝炎(乙肝)中的临床意义。方法采用高效液相色谱法(HPLC)测定60例慢性乙肝患者(乙肝患者组)和41例正常人(正常对照组)血浆中色氨酸和犬尿氨酸含量,并对两组数据进行比较。结果正常对照组Trp含量为(76.99±10.88)μmol/L,Kyn含量为(2.16±0.32)μmol/L,Kyn/Trp为0.028±0.003;乙肝患者组Trp含量为(68.04±8.52)μmol/L,Kyn含量为(2.18±0.26)μmol/L,Kyn/Trp为0.032±0.004。乙肝患者组血浆Trp含量明显低于正常对照组(P<0.01),Kyn/Trp明显高于正常对照组(P<0.01),两组Kyn含量比较差异无统计学意义(P>0.05)。结论慢性乙肝患者色氨酸代谢发生异常,检测患者血浆中Trp和Kyn有重要的临床意义。     

吲哚胺-2,3-双加氧酶活性与非小细胞肺癌进展的关系

目的 吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)及其介导的色氨酸(tryptophan,Trp)-犬尿氨酸(kynurenine,Kyn)代谢途径参与了肿瘤免疫逃逸.本研究探讨IDO活性与非小细胞肺癌(NSCLC)临床特征的关系.方法 利用高效液相色谱技术同期检测87例NSCLC患者、41例肺良性肿瘤患者及30例健康对照组血清中Kyn 与Trp浓度,IDO活性以血清中两者比值Kyn/Trp表示.结果 与30例健康对照组相比,NSCLC患者血清Trp水平明显降低 [(36.91 ±9.26)μmol/L vs(44.37±10.18)μmol/L,P＜0.01],其降低程度在NSCLC患者T4 [(35.51±9.12)μmol/L]及N3[(34.58±10.53)μmol/L] 亚组中尤为明显.但肺良性肿瘤组与健康对照组比较,Trp水平无明显差异.NSCLC组代表IDO活性的Kyn/Trp比值较健康对照组明显升高[(49.62 ±14.76)vs(30.32 ±11.43),P＜0.01],其差异有统计学意义.在NSCLC各亚组中,M1亚组的IDO活性程度最高,达到了(55.06±18.54).NSCLC组IDO活性在肿瘤切除术后1周时明显减弱.结论 NSCLC患者体内IDO活性明显增强,且与临床分期呈正相关.提示IDO可能参与了NSCLC的进展,代表其活性的Kyn/Trp 比值可作为判断NSCLC是否有远处转移的潜在指标.     

液相色谱-电喷雾串联质谱法分析大肠癌患者尿液色氨酸及其代谢产物

目的：通过液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）测定尿液色氨酸及其代谢产物浓度,探讨大肠癌患者体内色氨酸代谢变化及其与机体免疫之间的关系。方法通过 LC-ESI-MS/MS法测定大肠癌患者和健康者（对照组）尿液色氨酸代谢物浓度,并通过犬尿氨酸/色氨酸比值来评估代谢过程中的酶活性。结果大肠癌患者尿液犬尿氨酸、3-羟基犬尿氨酸（HK）、羟基氨茴酰酸（HAA）、吲哚-3-乳酸（ILA）、吲哚-3-丙酸（IPA）、邻氨基苯甲酸（AA）浓度显著升高（P＜0．05,P＜0．01）;而大肠癌患者尿液犬尿烯酸浓度较对照组显著下降（P＜0．05）;大肠癌患者犬尿氨酸/色氨酸比值较对照组显著提高（P＜0．01）。结论大肠癌患者尿液色氨酸代谢发生紊乱,色氨酸代谢物检测或可为大肠癌发病机制、预测和防治研究奠定基础。     

IDO/TTS介导的色氨酸代谢途径在免疫性血小板减少症患者发病及治疗中的作用

目的 探讨IDO/TTS介导的色氨酸代谢途径在免疫性血小板减少症患者的发病和治疗中的作用。 方法 选取2015年7月~2017年8月我院收治的36例免疫性血小板减少症患者和36例来我院体检的健康人为研究对象，分别为观察组和对照组，观察组给予地塞米松40 mg/d进行治疗，对照组不采取任何处理。通过液相-质谱分析仪检测血浆中犬尿氨酸（Kyn）和色氨酸（Trp）的浓度；通过流式细胞仪检测T淋巴细胞中IDO和TTS的表达。 结果 与对照组相比，治疗前免疫性血小板减少症患者血浆中Kyn、Trp、Kyn/Trp的表达量较高，IDO的表达量较低，而TTS的表达量则明显较高，差异有统计学意义（P<0.05）；观察组经有效治疗后，患者血浆中Trp的浓度降低，而Kyn浓度及Kyn/Trp比值则明显升高，差异有统计学意义（P<0.05）；患者IDO表达量明显上升，而TTS的表达量则明显下降，差异有统计学意义（P<0.05）。 结论 地塞米松对免疫性血小板减少症患者疗效显著，主要通过IDO/TTS介导的色氨酸代谢途径发挥作用，临床可以通过监测IDO、TTS的水平指导其治疗。     

基于HPLC的标准加入法同时测定小鼠血清中犬尿氨酸和色氨酸

 目的 建立一种同时测定小鼠血清中犬尿氨酸和色氨酸浓度的高效液相色谱(HPLC)法,用于评价小鼠给予IDO抑制剂后体内IDO活性。方法 血清用6%高氯酸沉淀蛋白法处理,离心后取上清进样,采用标准加入法定量。选用ThermoHypersil GOLD色谱柱(250 mm×4.6 mm,5 μm)分离,流动相为乙腈-醋酸钠(15 mmol/L,冰醋酸调pH至 4.0),流速1.1 mL/min,柱温30 ℃,犬尿氨酸和色氨酸的紫外检测波长分别为360 nm和280 nm。结果 犬尿氨酸保留时间为6.10 min,线性范围为0.25～10 μmol/L,检测限为0.09 μmol/L;色氨酸保留时间10.12 min,线性范围为25～1 000 μmol/L,检测限为0.25 μmol/L。精密度、回收率及稳定性等均符合生物样品定量分析方法验证指导原则的要求,并成功应用于62只小鼠血清犬尿氨酸和色氨酸浓度的测定。结论 建立了一种简单、快速、准确的HPLC分析方法,采用标准加入定量方式,可同时测定小鼠血清中犬尿氨酸和色氨酸血药浓度,并成功应用于实际生物样本的测定。     

高效液相色谱法测定人血清中5-HT含量

目的:建立以高效液相色谱法测定人血清中5-羟色胺(5-HT)含量的方法。方法:色谱柱为SinoChromODS-BP柱(250mm×4.6mmID,5μm0.05,大连依利特),流动相为0.05mol/L KH2PO4:甲醇=95∶5(磷酸调pH至4.0),柱温为40℃,检测波长为275nm,流速为1.0ml/min。结果:5-HT检测进样量在0.01μg~1.6μg范围内线性关系良好(r=0.9999),平均回收率为84.4%(RSD=1.096%,n=5)。结论:本方法调整了流动相的比例,调整了5-HT的保留时间。与文献报道方法相比,减少了杂质峰的干扰,更加简便、准确、重复性好,故可用于血清中5-HT含量测定。     

色氨酸-犬尿氨酸代谢通路与神经退行性疾病及中医药干预作用的研究进展

色氨酸代谢是一个可高度调控的生理过程，犬尿氨酸通路是色氨酸代谢的主要途径，色氨酸-犬尿氨酸代谢通路涉及一系列酶促反应，代谢生成的多种生物活性物质统称为“犬尿氨酸能物质”。越来越多的研究表明，这些犬尿氨酸能物质与很多神经系统疾病的发生、发展有着密切关联，异常的色氨酸-犬尿酸代谢与诸如帕金森氏病、亨廷顿病和阿尔茨海默症等神经退行性疾病认知功能低下有关。文章就色氨酸-犬尿氨酸通路的代谢、生成的神经生物学活性物质、相关的神经系统疾病以及以此为靶点的中医药干预作用进行综述，为研究和治疗相应神经系统疾病提供一定策略与参考。     

维持性血液透析患者精神心理状况分析及其与血清色氨酸水平的相关性研究

目的研究维持性血液透析患者精神心理状况及其与血清色氨酸水平的相关性。方法选取我院2015年3月~2016年12月122例进行维持性血液透析的患者及115例健康志愿者为研究对象,将122例进行维持性血液透析的患者纳入研究组,115例健康志愿者纳入对照组。检测两组焦虑和抑郁等心理状况及血清色氨酸水平。结果 (1)研究组患者焦虑阳性发生率和抑郁阳性发生率均高于对照组(P0.05)。(2)抑郁组患者IL-6、犬尿氨酸高于非抑郁组患者,抑郁组患者Trp、HGB低于非抑郁组患者(P0.05)。(3)焦虑组患者Scr、BUN高于非焦虑组患者,焦虑组患者血钙、血磷低于非焦虑组患者(P0.05)。(4)抑郁与IL-6、犬尿氨酸呈显著正相关性(r=0.351,P=0.021;r=0.256,P=0.002),焦虑与Scr、BUN呈显著正相关性(r=0.152,P=0.034;r=0.689,P=0.016)。结论炎性因子诱导使维持性血液透析患者色氨酸代谢成犬尿氨酸,中枢内5-HT水平降低,促使患者出现抑郁和焦虑等负面心理情绪,且抑郁与IL-6、犬尿氨酸水平呈显著正相关性,焦虑与Scr、BUN水平呈显著正相关性。     

构建LPS诱导的急性炎症小鼠模型用于IDO1抑制剂药效动力学评价

 目的 构建急性炎症小鼠模型用于吲哚胺2, 3-双加氧酶1（indoleamine 2,3-dioxygenase 1,IDO1）抑制剂的药效动力学评价。方法 采用尾静脉注射或腹腔注射给予小鼠脂多糖（lipopolysaccharide,LPS）,构建IDO1活性上调的急性炎症小鼠模型。模型小鼠灌胃给予IDO1抑制剂L-1-甲基色氨酸（L-1-methyl-tryptophan,L-1-MT）或TQ016,给药后不同时间点取血,HPLC检测小鼠血清中色氨酸（tryptophan,Trp）和犬尿氨酸（kynurenine,Kyn）含量,计算Kyn/Trp比值。采用IDO1活性上调引起的Trp浓度降低、Kyn浓度升高及Kyn/Trp比值增大,来评价IDO1抑制剂L-1-MT和TQ016对急性炎症小鼠血清中上调的IDO1活性的抑制效果。结果 尾静脉注射0.66 mg/kg LPS或腹腔注射5 mg/kg LPS均显著上调小鼠血清中IDO1活性。100 mg/kg L-1-MT在灌胃给药4 h后显著抑制模型小鼠血清中上调的IDO1活性。100 mg/kg TQ016在灌胃给药1、6、12、24和30 h后均显著抑制模型小鼠血清中上调的IDO1活性。TQ016呈剂量依赖性抑制模型小鼠血清中上调的IDO1活性。结论 LPS诱导的急性炎症小鼠模型适用于IDO1抑制剂的药效动力学评价。IDO1抑制剂TQ016具有良好的IDO1活性抑制效果。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.