乙肝病毒携带者外周血单个核细胞培养上清液HBV-DNA定量检测意义

目的　探讨慢性无症状乙肝病毒携带者外周血单个核细胞 (PBMC)与HBV -DNA定量的临床关系及意义。方法　采用慢性乙肝病毒HBeAg阳性携带者PBMC体外刺激诱导培养 ,荧光定量PCR(FQ -PCR)法测定上清液HBV -DNA ,ELISA法检测白细胞介素 10、12 (IL -10、12 )与γ 干扰素 (γ IFN)。同期检测肝功能 ,血清HBV -DNA定性试验。结果　PBMC培养后于上清液中以FQ -PCR法检出HBV -DNA。 >5 0 0 0考贝 /ml组中 ,IL -12及γ IFN均明显高于正常对照组。重症肝炎PBMC上清液FQ -PCR测定HBV -DNA阳性率明显高于同期血清PCR定性试验阳性率。PBMC上清液FQ -PCR测定HBV -DNA阳性率与血清HBeAg转换的关系同血清HBV -DNA定性试验阳性率与其关系相仿。与肝功能ALT及TBiL程度无明显相关。结论　HBeAg携带者PBMC培养液HBV -DNA高复制状态时IL -12及γ IFN分泌增强。重症肝炎PBMC上清液HBV -DNAFQ -PCR测定阳性率高于同期血清PCR定性实验     

Polyunsaturated Fatty Acids and Their Metabolites in Hyperemesis Gravidarum

Polyunsaturated fatty acids (PUFAs) have been studied in relation to pregnancy. However, there is limited knowledge on PUFAs and their metabolites in relation to hyperemesis gravidarum (HG), a pregnancy complication associated with nutritional deficiencies and excessive vomiting. In order to survey the field, a systematic review of the literature was performed, which also included nausea and vomiting of pregnancy (NVP) due to its close relationship with HG. In the very few published studies found, the main subjects of the research concerned free fatty acids (four records), lipid profiles (three records), and bioactive lipids (one article about prostaglandin E₂ and one about endocannabinoids). The authors of these studies concluded that, although no cause-and-effect relationship can be established, HG is linked to increased sympathetic responsiveness, thermogenic activity and metabolic rate. In addition, NVP is linked to a metabolic perturbance (which lasts throughout pregnancy). The low number of retrieved records underlines the need for more research in the area of PUFAs and HG, especially with regard to the underlying mechanism for the detected effects, potentially involving growth differentiation factor 15 (GDF15) since evidence for GDF15 regulation of lipid metabolism and the role for GDF15 and its receptor in nausea and vomiting is emerging.     

脂肪乳剂对健康人外周血单个核细胞磷脂脂肪酸组成的影响

目的探讨脂肪乳剂对健康人免疫细胞脂肪酸组成的影响。方法采用毛细管气相色谱法检测8名健康志愿者连续7天外周静脉输注脂肪乳剂(20%intralipid)后,外周血单个核细胞磷脂脂肪酸谱的变化。结果静脉输注脂肪乳剂对健康人外周血单个核细胞数量、肝功能和血脂无影响,软脂酸(C16∶0)、油酸(C18∶1N9)比例在单个核细胞磷脂酰乙醇胺(PE)中明显增加(P<0.05,P<0.01);硬脂酸(C18∶0)减少(P<0.05);亚麻酸(C18∶3N3)、软油酸(C16∶1N7)在磷脂酰胆碱(PC)中明显增加(P<0.05),而其余脂肪酸均未发生明显改变。饱和脂肪酸与不饱和脂肪酸的比值(S/U)和脂肪酸的不饱和指数(UI)在单个核细胞PE、PC中均保持不变。结论脂肪乳剂不可能通过影响免疫细胞磷脂脂肪酸组成的途径对免疫功能产生影响。     

Fingertip Whole Blood as an Indicator of Omega-3 Long-Chain Polyunsaturated Fatty Acid Changes during Dose-Response Supplementation in Women: Comparison with Plasma and Erythrocyte Fatty Acids

The sensitivity of fingertip whole blood to reflect habitual dietary and dose-dependent supplemental omega-3 long-chain polyunsaturated fatty acid (n-3 LCPUFA) intake in premenopausal women was compared to that of venous erythrocytes and plasma fatty acids. Samples were obtained from women in a randomised, double-blind, placebo-controlled trial in which premenopausal women (n = 53) were supplemented with DHA-rich tuna oil capsules and/or placebo (Sunola oil) capsules (6 capsules per day) for 8 weeks to achieve doses of either 0, 0.35, 0.7 or 1.05 g/day n-3 LCPUFA. All blood biomarkers were very similar in their ability to reflect dietary n-3 LCPUFA intake (r = 0.38–0.46 for EPA and DHA intake), and in their dose-dependent increases in n-3 LCPUFA levels after supplementation (R² = 0.41–0.51 for dose effect on biomarker EPA and DHA levels (mol %)). Fingertip whole blood is an effective alternative to erythrocytes and plasma as a biomarker n-3 LCPUFA intake in premenopausal women.     

Yellow Gentian Root Extract Provokes Concentration- and Time-dependent Response in Peripheral Blood Mononuclear Cells

Yellow gentian (Gentiana lutea L.), a medicinal plant widely used in traditional medicine, displays multiple biological effects, ranging from beneficial to toxic. Since many promising applications have been reported so far, our aim was to evaluate its potential concentration- and time- dependent cytotoxic and genotoxic effects in vitro. To that end we exposed human peripheral blood mononuclear cells to 0.5, 1, and 2 mg/mL of yellow gentian root extract (YGRE) to determine its effects on oxidative stress parameters [pro/antioxidant balance (PAB) and lipid peroxidation], DNA damage (alkaline comet assay and chromosome aberrations), and cell viability (trypan blue exclusion test). Cell viability decreased with increasing concentrations and treatment duration. Only the lowest YGRE concentration (0.5 mg/mL) increased oxidative stress but produced minor DNA damage and cytotoxicity. At higher concentrations, redox parameters returned to near control values. The percentage of chromosome aberrations and percentage of DNA in the comet tail increased with increased YGRE concentration after 48 h and declined after 72 h of treatment. This points to the activation of DNA repair mechanism (homologous recombination), evidenced by the formation of chromosomal radial figures after 72 h of treatment with the highest YGRE concentration of 2 mg/mL. Our results suggest that YGRE, despite induction of cytotoxic and genotoxic effects, activates cell repair mechanisms that counter oxidative and DNA lesions and induce cell death in highly damaged cells. Therefore, observed protective effects of yellow gentian after longer exposure could be a result of activated repair and removal of cells with irreparable damage.Key words: cytotoxicity, genotoxicity, Gentiana lutea L., homologous recombination, redox parameters     

荧光定量PCR检测慢性乙肝患者外周血单个核细胞乙肝病毒 DNA

目的探讨定量检测慢性乙肝患者外周血单个核细胞(PBMC)乙型肝炎病毒DNA(HBV-DNA)的临床意义.方法采用荧光定量PCR检测PBMC及血清中HBV-DNA含量.结果 49例慢性乙肝患者PBMC及血清中HBV-DNA阳性率分别为44.9%(22/49),51.0%(25/49),两者阳性率无统计学意义的差异(P＞0.05),具有一致性;血清HBV-DNA高水平组PBMC的HBV-DNA含量与低水平组比较,两者有统计学意义差异(P＜0.01);血清HBV- DNA高水平组PBMC的HBV-DNA阳性率(100%)与血清低水平组PBMC的HBV-DNA阳性率(42.9%)比较,两者有统计学意义的差异(P＜0.05);在24例血清HBV-DNA阴性患者中,发现5例PBMC HBV-DNA阳性.结论 PBMC的HBV-DNA检测可反映病毒在体内的复制程度,是对慢乙肝患者血清HBV-DNA检测有意义的补充,有助于临床上对慢乙肝患者血清病毒复制状态的了解及指导治疗用药.     

正常孕妇妊娠不同阶段外周血巨噬细胞移动抑制因子基因表达水平及其血清中含量的变化

目的 检测正常孕妇妊娠不同阶段外周血单个核细胞(PBMC)巨噬细胞移动因子(MIF)基因表达水平及血清中MIF含量的变化,为揭示妊娠期间的调控机制提供实验依据.方法 42名妊娠不同阶段的正常孕妇来自天津市中医药大学第一附属医院,年龄26～36岁,平均(31.0±3.0)岁,其中妊娠早期18例,孕龄(8.71±1.35)周;妊娠中期9例,孕龄(21.35±3.37)周;妊娠晚期15例,孕龄(33.01±2.32)周.9例非妊娠对照组来自天津市血液中心健康女性志愿者,年龄(26.5±3.0)岁.用密度梯度离心法分离正常孕妇PBMC,调整细胞浓度为1×106个,提取RNA,反转录(RT)合成cDNA,用荧光定量聚合酶链反应(PCR)方法检测MIF mRNA表达水平,用酶联免疫吸附(ELISA)法检测孕妇血清中MIF含量.结果 妊娠早期、中期、晚期PBMC MIF基因表达水平分别为2.15±0.47、2.07±0.39、2.18±0.54,与非妊娠对照组(0.68±0.12)比较,均有明显升高,差异有统计学意义(P＜0.05).妊娠早期、中期、晚期血清中MIF含量分别为(15.25±1.72)、(16.12±1.99)、(15.55±1.82)ng/ml,与非妊娠对照组[(5.53±1.77)ng/ml]比较,均有明显升高,差异有统计学意义(P＜0.05).不同妊娠时间比较,MIFmRNA表达水平和MIF含量的差异均无统计学意义(P＞0.05).结论 MIF的基因表达和含量在整个妊娠过程中都维持在较高水平,提示MIF参与了整个妊娠至分娩.     

血浆脂肪酸和食物频率法评估脂肪摄入的相关性研究

<正>食物频率法(FFQ)是常用的膳食调查方法之一,它经过几十年的研究发展可分为定性、定量和半定量食物频率法。食物频率法优点很多,但其缺点是存在回忆偏倚,因此需要对其准确度进行评价,国外很多研究显示食物频率法对各种营养素评价有优劣之分,可较好的反映体内某些营养素,但对于另外一些营养素又不适用。Hodson等的研究表明[1]血浆脂肪酸是评价膳食脂肪酸的良好指标。本研究以α-亚麻酸、亚油酸和硬脂酸作为生物标志物,比较膳食脂肪酸与血浆脂肪     

Omega-3 Polyunsaturated Fatty Acids Trigger Cell Cycle Arrest and Induce Apoptosis in Human Neuroblastoma LA-N-1 Cells

Omega-3 (n-3) fatty acids are dietary long-chain fatty acids with an array of health benefits. Previous research has demonstrated the growth-inhibitory effect of n-3 fatty acids on different cancer cell lines in vitro, yet their anti-tumor effects and underlying action mechanisms on human neuroblastoma LA-N-1 cells have not yet been reported. In this study, we showed that docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) exhibited time- and concentration-dependent anti-proliferative effect on the human neuroblastoma LA-N-1 cells, but had minimal cytotoxicity on the normal or non-tumorigenic cells, as measured by MTT reduction assay. Mechanistic studies indicated that DHA and EPA triggered G₀/G₁ cell cycle arrest in LA-N-1 cells, as detected by flow cytometry, which was accompanied by a decrease in the expression of CDK2 and cyclin E proteins. Moreover, DHA and EPA could also induce apoptosis in LA-N-1 cells as revealed by an increase in DNA fragmentation, phosphatidylserine externalization and mitochondrial membrane depolarization. Up-regulation of Bax, activated caspase-3 and caspase-9 proteins, and down-regulation of Bcl-X_L protein, might account for the occurrence of apoptotic events. Collectively, our results suggest that the growth-inhibitory effect of DHA and EPA on LA-N-1 cells might be mediated, at least in part, via triggering of cell cycle arrest and apoptosis. Therefore, DHA and EPA are potential anti-cancer agents which might be used for the adjuvant therapy or combination therapy with the conventional anti-cancer drugs for the treatment of some forms of human neuroblastoma with minimal toxicity.     

Association Between Low Colonic Short-Chain Fatty Acids and High Bile Acids in High Colon Cancer Risk Populations

We propose that the influence of diet on colon cancer risk is mediated by the microbiota. To investigate how dietary fat influences risk, we compared the colonic contents of 12 adult high-risk African Americans (AAs) and 10 Caucasian Americans (CAs) who consumed a high-fat diet (123 ± 11 g/d and 129 ± 17 g/d, respectively) to 13 native Africans (NAs) who subsisted on a low-fat (38 ± 3.0 g/d) diet, all aged 50–60 yr. The colonic bile acids were measured by LC-MS and the short-chain fatty acids (SCFAs) by GC. The chief secondary colonic bile acids, deoxycholic acid and lithocholic acid, were correlated with fat intake and similar between AAs and CAs, but 3–4 times higher than in AAs (p < 0.05). The major SCFAs were lower in AAs (p < 0.001) and CAs (p < 0.001) compared to AAs, but conversely, the branched chain fatty acids (BFCA) were higher. Our results suggest that the higher risk of colon cancer in Americans may be partly explained by their high-fat and high-protein, low complex carbohydrate diet, which produces colonic residues that promote microbes to produce potentially carcinogenic secondary bile acids and less antineoplastic SCFAs. The role of BCFA in colonic carcinogenesis deserves further study.     

多重PCR法检测外周血单个核细胞中的HBVDNA与HBVcccDNA

目的建立多重聚合酶链反应法(Multiplex Polymerase chain reaction，M—PCR)，揭示慢性乙肝患者外周血单个核细胞(PBMC)中HBVDNA的存在状况。方法采用一种把一套扩增乙肝病毒基因组DNA(HBV genome DNA)的引物和一套特异性扩增乙肝病毒共价闭合环状DNA(HBVcccDNA)的引物掺入到一个体系中的PCR法，检测PBMC中的HBVDNA核酸分子。结果PCR法可使处于同一PBMC中的总HBVDNA和HBVcccDNA同时分别良好的扩增出来；30例慢性乙肝患者的PBMC中，总HBVDNA与HBVcccDNA同耐检出者23例，检出率76．6％；检出HBVcccDNA者占检出总HBVDNA者的82．1％。结论PBMC中的HBVDNA部分参与复制。成功的建立了能同时检测HBV基因组DNA与HBVcccDNA的M—PCR方法。     

Preventive Activity of Olive Oil Phenolic Compounds on Alkene Epoxides Induced Oxidative DNA Damage on Human Peripheral Blood Mononuclear Cells

The aim of this study was to investigate the ability of epoxides of styrene (styrene-7,8-oxide; SO) and 1,3-butadiene (3,4-epoxy-1-butene; 1,2:3,4:-diepoxybutane) to cause oxidative stress and oxidative DNA damage on human peripheral blood mononuclear cells (PBMCs) and whether a complex mixture of olive oil phenols (OOPE) could prevent these effects. The DNA damage was measured by the single-cell gel electrophoresis (SCGE; comet assay). We found that the DNA damage induced by alkene epoxides could be prevented by N-acetyl-cysteine (10 mM) and catalase (100 U/ml). Alkene epoxides caused a significant (P < 0.05) increase of both peroxide concentration in extra- and intracellular environment and formamidopyrimidine DNA glycosylase (FPG)- and Endonuclease III (ENDO III)-sensitive sites in PBMCs, demonstrating the presence of oxidized bases. OOPE (1 μg of total phenols/ml) was able to prevent the alkene epoxide induced DNA damage both after 2 and 24 h of incubation. In addition, OOPE completely inhibited the SO-induced intracellular peroxide accumulation in PBMCs and prevented the oxidative DNA damage induced by SO, as evidenced by the disappearance of both FPG- and ENDO III-sensitive sites. This is the first study demonstrating the ability of OOPE to prevent the DNA damage induced by alkene epoxides providing additional information about the chemopreventive properties of olive oil.     

溃疡性结肠炎患者外周血单个核细胞IL-13mRNA表达的研究

目的探讨溃疡性结肠炎(UC)患者外周血单个核细胞(PBMCs)白介13(IL-13)mRNA表达与血清C反应蛋白(C-RP)的关系。方法对21例溃疡性结肠炎(UC)活动期患者、10例缓解期患者及20例健康人,采用RT-PCR法和图像分析半定量法检测PBMCs中IL-13mRNA的表达水平及血清C反应蛋白(CRP)浓度。结果活动期UC患者IL-13mRNA的表达水平(6.45±1.23)明显低于缓解期UC患者(14.72±2.12)和健康人(15.17±2.38)(t=2.13,2.24,P<0.05),而缓解期UC患者与健康人比较无明显降低(t=1.63,P>0.05)。活动期UC患者IL-13mRNA的表达水平(6.45±1.23)与血清C-RP浓度(8.36±3.16mg/L)呈负相关(r=-0.589,P<0.05)。结论活动期UC患者PBMCs中IL-13mRNA表达水平显著降低,炎症活动程度与IL-13基因转录降低有关。     

正常孕妇外周血单个核细胞IL-8基因表达水平及血清中IL-8含量

摘要：目的检测妊娠不同阶段正常孕妇外周血单个核细胞（PBMC）IL-8基因表达水平及血清中IL-8含量。方法用密度梯度离心法分离正常孕妇PBMC,调整细胞浓度为1×10^6,提取RNA,逆转录合成cDNA,用荧光定量PCR方法检测IL-8mRNA表达水平。同时取正常孕妇外周血,分离血清,用ELISA和化学发光法检测孕妇血清中IL-8及人绒毛膜促性腺激素（HCG）含量。结果在妊娠初期,IL-8的表达（0.029±0.010）和分泌[（49．42±6．68）ng／ml]随着HCG含量的增加达到最高,在妊娠中期和晚期,IL-8的表达（中期0．004±0．001,晚期0．005±0.001和分泌[中期（8．17±3．09）ng/ml,晚期（7．98±3．05）ng/ml]又降到较低的水平。结论IL-8在妊娠初期的表达水平明显高于妊娠中期和晚期,提示,在HCG作用下IL-8参与了妊娠的启动。     

γ射线外照射诱发DNA损伤的研究

[目的] 研究γ射线对小鼠外周血淋巴细胞DNA的损伤效应.[方法] 分别应用中性、碱性单细胞凝胶电泳技术检测不同剂量γ射线外照射诱导小鼠外周血淋巴细胞DNA的单链和双链断裂.[结果] 0、1.0、5.0、10.0、15.0、20.0、30.0、40.0、60.0 Gy γ射线照射小鼠外周血淋巴细胞后,因DNA双链断裂而发生迁移的细胞比率分别为3.9%、9.0%、28.0%、32.0%、44.0%、56.0%、64.0%、96.0%、96.0%,细胞DNA迁移长度分别为(23.42±1.32)μm、(30.50±2.14)μm、(37.43±0.75)μm、(47.87±1.50)μm、(54.33±0.33)μm、(60.72±1.93)μm、(71.66±0.68)μm、(74.00±1.86)μm、(66.41±3.68)μm.0、0.5、1.0、3.0、5.0、10.0 Gy γ射线照射小鼠外周血淋巴细胞后,因DNA单链断裂而发生迁移的细胞比率分别为1.80%、7.20%、8.10%、10.90%、23.68%、27.77%,细胞DNA迁移长度分别为(21.59±0.49)μm、(25.58±1.13)μm、(27.09±0.5)μm、(27.18±0.66)μm、(36.25±1.15)μm、(53.28±0.13)μm.γ射线外照射对小鼠外周血淋巴细胞DNA具有明显的损伤作用,彗星细胞百分率显著增加,DNA电泳迁移,且迁移的距离与照射剂量之间呈良好的线性关系.[结论] 单细胞凝胶电泳不但能用于检测辐射所致离体细胞DNA单链的损伤效应,还能用来研究离体细胞DNA双链的辐射损伤和修复作用,并有望成为新一代生物剂量计.     

Association between Preoperative Long-Chain Polyunsaturated Fatty Acids and Oxidative Stress Immediately after Total Knee Arthroplasty: A Pilot Study

Quadriceps muscle atrophy following total knee arthroplasty (TKA) can be caused by tourniquet-induced ischemia–reperfusion (IR) injury, which is often accompanied by oxidative stress and inflammatory responses. n-3 long-chain polyunsaturated fatty acids (LCPUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exert antioxidant and anti-inflammatory effects against IR injury, whereas n-6 LCPUFAs, particularly arachidonic acid (AA), exhibit pro-inflammatory effects and promote IR injury. This study aimed to examine whether preoperative serum EPA + DHA levels and the (EPA + DHA)/AA ratio are associated with oxidative stress immediately after TKA. Fourteen eligible patients with knee osteoarthritis scheduled for unilateral TKA participated in this study. The levels of serum EPA, DHA, and AA were measured immediately before surgery. Derivatives of reactive oxygen metabolites (d-ROMs) were used as biomarkers for oxidative stress. The preoperative serum EPA + DHA levels and the (EPA + DHA)/AA ratio were found to be significantly negatively correlated with the serum d-ROM levels at 96 h after surgery, and the rate of increase in serum d-ROM levels between baseline and 96 h postoperatively. This study suggested the preoperative serum EPA + DHA levels and the (EPA + DHA)/AA ratio can be negatively associated with oxidative stress immediately after TKA.     

Sysmex XN-3000血液分析仪检测外周血有核红细胞的临床价值评估

目的:评估Sysmex XN-3000血液分析仪检测外周血有核红细胞的临床价值。方法:选取2017年1月~2018年7月在本院住院患者80例,对患者血液标本均进行Sysmex XN-3000血液分析仪检测及显微镜镜检法,观察检测结果。结果:将显微镜镜检法作为参照标准,Sysmex XN-3000检测外周血有核红细胞阴性28例,血片手工复检1例外周血有核红细胞阳性,Sysmex XN-3000检测外周血有核红细胞阳性52例,血片手工复检50例阳性,Sysmex XN-3000检测敏感性98.04%(50/51),特异性93.10%(27/29)。结论:Sysmex XN-3000血液分析仪检测外周血有核红细胞,操作简便快捷,具有较高的准确性。     

Long-Chain Polyunsaturated Fatty Acid Concentrations and Association with Weight Gain in Pregnancy

Lower concentrations of omega-3 (ω-3) and higher concentrations of omega-6 (ω-6) have been associated with excess weight in adults; however, the information on this relationship in pregnancy remains in its infancy. This study aimed to investigate the association between plasma levels of ω-3 and ω-6 long-chain polyunsaturated fatty acids (PUFAs) and weight gain during the gestational period. This is a prospective cohort study involving 185 pregnant women registered with the prenatal services of a municipality in the northeast of Brazil. The dosage of the serum concentration of fatty acids and the anthropometric measurements were carried out at the baseline, and the women’s weight information in the first, second, and third trimesters was collected from their pregnancy cards. Serum fatty acids were determined with the help of gas chromatography. The response variable of this study is the latent variable weight gain in pregnancy, derived from three variables: gestational weight in the first, second, and third trimesters. The main exposure was the plasma concentrations of PUFAs. Structural equation modeling was used for the data analysis. The mean age of the pregnant women was 26.74 years old (SD: 5.96 years). Most of the women had not completed high school (84%) and had a low income (70.86%). It was observed that the ω-3 PUFAs, represented by ALA plasm (alpha-linolenic acid), DHA (docosahexaenoic acid), and the EPA/ALA ratio (eicosapentaenoic acid to alpha-linolenic acid ratio), were negatively associated with the weight gain during pregnancy construct (−0.20, −0.12, and −0.14, respectively). Meanwhile, the PUFAs represented by the ratio between the ω-6 category acids ARA and LA (arachidonic acid and linoleic acid) had a direct and positive association (0.22) with that construct. Excess maternal weight gain was associated with ω-3 and ω-6 plasma levels. The women with the greatest gestational weight gain were the ones that presented the highest ARA/LA ratio (ω-6) and the lowest plasma concentrations of ALA, DHA, and EPA/ALA ratio (ω-3).     

美白祛斑类化妆品使用者尿汞与外周血细胞DNA损伤的关系

目的研究高含汞化妆品对人体外周血细胞DNA的损伤效应。方法根据化妆品中汞含量将49例使用美白祛斑类化妆品的女性分为汞超标组(1mg/kg,29人)和汞未超标组(≤1mg/kg,20人),年龄为21～57岁,使用时间为2个月～7年。测定尿汞含量;采用单细胞凝胶电泳技术检测外周血DNA损伤情况。结果尿汞值与彗星的头部面积、头部荧光强度、头部百分比、头部半径均呈负相关,与彗星尾部面积、尾部荧光强度、尾部百分比、尾长、尾矩、Oliver尾矩均呈正相关,差异有统计学意义(P0.01);而与全长无相关性(P0.05)。结论化妆品中的汞通过皮肤进入体内可引起汞蓄积,导致人体外周血细胞DNA受损。