Expression and significance of toll-like receptor 2,4 in peripheral blood mononuclear cells in patients with systemic inflammatory response syndrome

To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical study was done on 103 patients of which 65 were with SIRS.The mRNA expression of TLR2,4 were detected by RT-PCR;the expression of TNF-α and IL-6 were observed by ELISA;the correlation between TLR2,4 mRNA,the level of TNF-α and IL-6,and the clinical course was evaluated.Results TLR2 mRNA ,TNF-α and IL-6 were upregulated markedly on the first day of hospitalization,then decreased gradually;TLR2 mRNA maintained on high level till the 5th day.The expression of TLR2,4 mRNA was positive correlated with the level of TNF-α and IL-6,and the length of stay.TLR2,4 mRNA expression increased in patients with multiple organ failure.Conclusion In actue abdomen patients with SIRS,the expression of TLR2,4 of PBMC increased markedly,indicating its improtant role in the pathogenesis of SIRS.4 refs,2 figs,2 tabs.

Protection against hepatic ischemia/reperfusion injury via downregulation of toll-like receptor 2 expression by inhibition of Kupffer cell function

AIM： To elucidate the mechanism of liver protection by inhibition of Kupffer cells （KCs） function.METHODS： All the animals were randomly divided into three groups. Blockade group （gadolinium chloride solution （GdCl3） injection plus ischemia/reperfusion （I/R） injury）：GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group （saline solution injection plus I/R injury）： saline instead of GdCl3 as a control was injected as in the blockade group. Sham group： saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. Toll-like receptor2 （TLR2） was immunostained with a goat antimouse polydonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-α （TNF-α） and alanine aminotransferase （ALT）, an indicator of liver function.I/R injury via downregulation of theexpression of TLR2.RESULTS： Compared to non-blockade group, CD^68＋ cells significantly reduced in blockade group （OPTDI, optical density integral）： 32.97±10.55 vs 185.65±21.88, P〈0.01） and the liver function impairment was relieved partially （level of ALT： 435.89±178.37 U/L vs 890.21±272.91 U/L,P〈0.01）. The expression of TLR2 protein in blockade group significantly decreased compared to that in non- group（method of immunohistochemistry, OPDTI： 75.74±17.44 vs 170.58±25.14, P〈0.01; method of Western blot, A value： 125.89±15.49 vs 433.91±35.53, P〈0.02）. The latter correlated with the variation of CD68 staining （r = 0.745,P〈0.05）. Also the level of portal vein TNF-α decreased in blockade group compared to that in non-blockade group （84.45±14.73 ng/L vs 112.32±17.56 ng/L, P〈0.05）, butwas still higher than that in sham group （84.45±14.73 ng/L vs 6.07±5.33 ng/L, P〈0.01）.CONCLUSION： Inhibition of the function of KCs may protect liver against I/R injury via downregulation of the expression of TLR2.

Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads

The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.

Expression of Toll-like Receptor 4 in Neonatal Cord Blood Mononuclear Cells in Patients with Preeclampsia

The expression of Toll-like receptor 4 (TLR4) in neonatal cord blood mononuclear cells (MNCs) and serum TNF-α were investigated in order to explore the roles of TLR4 in the pathogenesis of preeclampsia.The study enrolled 27 patients suffering from preeclampsia (experimental group) and 21 normal pregnancy patients (control group).After MNCs were separated, the expression of TLR4 mRNA and protein was detected by using real-time quantitative PCR and Western blotting respectively, and the expression of TNF-α by using ELISA.The results showed the TLR4 mRNA level in cord blood MNCs (2-CT:0.07±0.17), TLR4 protein expression level (absorbance ratio:0.81%±0.15%) and TNF-α level (9.5±1.73 pg/mL) were all increased in experimental group as compared with control group with the differences being statistically significant (P【0.05).There was a positive correlation between the expression of TLR4 mRNA and TNF-α in both experimental group and control group (r=0.54 and 0.53, respectively, P【0.05).It was concluded that TLR4 expression in the experimental group of cord blood MNCs was increased and there was a positive correlation between the expression of TLR4 mRNA and TNF-α in both groups.TLR4-mediated release of inflammatory cytokines may be one of the important reasons leading to preeclampsia.

Upregulation of Toll-like Receptor 4 on T Cells in PBMCs Is Associated with Disease Aggravation of HBV-related Acute-on-chronic Liver Failure

Summary： Immune-mediated inflammatory injury is an important feature of the disease aggravation of hepatitis B virus-related acute-on-chronic liver failure （ACLF）. Toll-like receptors （TLRs） have been shown previously to play a pivotal role in the activation of innate immunity. The purpose of this study was.to characterize the TLR4 expression in peripheral blood mononuclear cells （PBMCs） of ACLF pa- tients and its possible role in the disease aggravation. Twelve healthy subjects, 15 chronic HBV-infected （CHB） patients and 15 ACLF patients were enrolled in this study. The TLR4 expression in PBMCs and T cells of all subjects was examined by real-time PCR and flow cytometry. The correlation of TLR4 ex- pression on T cells with the markers of disease aggravation was evaluated in ACLF patients. The ability of TLR4 ligands stimulation to induce inflammatory cytokine production in ACLF patients was ana- lyzed by flow cytometry. The results showed that TLR4 mRNA level was upregulated in PBMCs of ACLF patients compared to that in the healthy subjects and the CHB patients. Specifically, the expres- sion of TLR4 on CD4＋ and CD8＋ T cells of PBMCs was significantly increased in ACLF patients. The TLR4 levels on CD4＋ and CD8＋T cells were positively correlated with serum total bilirubin （TBIL）, direct bilirubin （DBIL）, international normalized ratio （INR） levels and white blood cells （WBCs）, and negatively correlated with serum albumin （ALB） levels in the HBV-infected patients, indicating TLR4 pathway may play a role in the disease aggravation of ACLF. In vitro TLR4 ligand stimulation on PBMCs of ACLF patients induced a strong TNF-α production by CD4＋ T cells, which was also posi- tively correlated with the serum markers for liver injury severity. It was concluded that TLR4 expression is upregulated on T cells in PBMCs, which is associated with the aggravation of ACLF.

Study of a Quantitative Fluorescence Method for Interleukin 2 Receptor of Mononuclear Cell in Normal Human Peripheral Blood

A quantitative method using fluorescence spectro-photometer to detect theinterleukin 2 reocptor （IL 2R） on the surface of mononuclear cells in normal humanperipheral blood is described. For expression of IL 2R, the optimum conoentration of PHA andCon A were 100 μg/ml and 10 μg/ml respectively. The PHA induced effect on mononuclear cellswas better than Con A induced one. The peak value of IL 2R cxpression was at the 24th h afterstimulation The optimum dilution of the first antibody （anti-Tac） was 1: 50, while the dilution ofthe second antibody（fluorescein conjugated rabbit anti-mousc IgG） was 1: 32 Their optimumincubation period was 20 and 10 min repectively. The concentration of fluorcscein-conjugatedantibody per 1×106 mononuclear cells from the same normal human peripheral blood was5.22±0.45×10-10 mol, and the coefficient of variation was 8. 6%. The concentration offluorescein-conjugated antibody per 1×106PBMC from five healthy individuals was 4. 8±0. 97×10-10 mol Therefore this assay system is stable and quite reproducible.

缺血性脑卒中患者TLR2 TLR4与血小板神经功能的关系

目的探究缺血性脑卒中患者外周血单核细胞(PBMC)中Toll样受体(TLR)2、TLR4表达水平与血小板活化及神经功能的关系。方法选取2017年5月至2020年6月邢台医学高等专科学校第二附属医院收治的134例缺血性脑卒中患者为研究对象(研究组),同期140例健康体检者为对照(对照组),收集两组对象一般资料。采用实时荧光定量PCR(RT-qPCR)检测两组对象PBMC中TLR2、TLR4 mRNA水平;利用酶联免疫吸附试验(ELISA)法检测两组对象血浆中血小板活化因子(PAF)、血小板活化标志物α颗粒膜糖蛋白140(GMP-140)、β-血小板球蛋白(β-TG)、血小板第4因子(PF4)水平;采用美国国立卫生院卒中量表(NIHSS)评分评估研究组患者神经功能;Spearman分析研究组患者PBMC中TLR2、TLR4与糖尿病史、高血压史的相关性;Pearson分析研究组患者PBMC中TLR2、TLR4与低密度脂蛋白胆固醇(LDL-C)、空腹血糖(FBG)、高密度脂蛋白胆固醇(HDL-C)水平,血浆中PAF、GMP-140、β-TG、PF4水平,NIHSS评分的相关性。结果与对照组相比,研究组糖尿病史、高血压史比例,LDL-C、FBG水平,PBMC中TLR2、TLR4水平,血浆中PAF、GMP-140、β-TG、PF4水平升高(P<0.05),HDL-C水平降低(P<0.05)。研究组NIHSS评分为(14.59±7.48)分。相关性分析发现,PBMC中TLR2、TLR4与PAF、β-TG、PF4、NIHSS评分,TLR4与LDL-C、FBG、GMP-140呈正相关(P<0.05)。结论缺血性脑卒中患者PBMC中TLR2、TLR4表达水平升高,其与血小板活化及神经功能评分具有一定相关性。

Co-regulation of Dectin-1 and TLR2 in inflammatory response of human corneal epithelial cells induced by Aspergillus fumigates

AIM： To investigate the co-regulation of dendritic cell- associated C-type lectin-1 （Dectin-1）, Toll-like receptor 2 （TLR2）, and relative chemotactic factors in the Telomease- immortalized human corneal epithelial （THCE） cells after exposure to ,Aspergillus fumigatus （Af） hyphae. METHODS： The normal THCE cells were investigated as control. After cultured in vitro with Af hyphae, with or without laminarin and anti-TLR2 antibody for 4, 8, 16 and 24h, THCE cells were harvested. The expression of Dectin-1, TLR2, CXCL1 and CXCL8 mRNA were measured by real-time quantitative polymerase chain reaction at the stimulation of 4, 8 and 16h separately. The protein expression of Dectin-1 and TLR2 were analyzed at 8, 16, and 24h by Western blot. ~ RESULTS： The mRNA CXCL8 increased in THCE expression of CXCL1 and cells after stimulated by Af hyphae. The stimulatory effects on these inflammatory chemokines were shown in a dose-dependent manner and reached the peak at 8h. Af hyphae significantly stimulated the production of Dectin-1 and TLR2 in THCE cells at both mRNA and protein levels. The protein of Dectin-1 and TLR2 gradually increased till 16h. While pretreated with laminarin （a Dectin-1 inhibitor）, the expression of TLR2, CXCL1 and CXCL8 all decreased dramatically at the peak point, interestingly, when pretreated with TLR2 neutralizing antibody, the expression of Dectin -1, CXCL1 and CXCL8 also decreased dramatically at the peak point. CONCLUSION： These findings suggest that Dectin-1 and TLR2 co-regulated with each other after treated with inactive Af hyphae in the THCE cells, and they contribute together to the inflammatory responses by induction of chemokines CXCL1 and CXCL8.

不同肝组织炎症分级和纤维化分期的自身免疫性肝炎患者外周血单个核细胞TLR2和CTLA-4水平变化研究

目的 调查不同肝组织炎症分级和纤维化分期的自身免疫性肝炎(AIH)患者外周血单个核细胞(PBMCs)Toll样受体2(TLR2)和细胞毒性T淋巴细胞相关抗原4(CTLA-4)水平变化。方法 2018年6月~2022年6月我院诊治的47例AIH患者和同期50例健康人,所有AIH患者接受肝活检。抽取外周血,分离PBMCs,采用PCR法检测PBMCs TLR2和CTLA-4 mRNA水平,使用流式细胞仪检测TLR2和CTLA-4阳性的PBMCs百分比。结果 AIH患者PBMCs TLR2 mRNA水平和细胞表面TLR2阳性的PBMCs百分比分别为(1.7±0.4)和(34.2±7.9)%,均显著高于健康人【分别为(0.9±0.2)和(21.4±3.5)%,P<0.05】,而PBMCs CTLA-4 mRNA水平和细胞表面CTLA-4阳性的PBMCs百分比分别为(0.5±0.2)和(2.3±0.8)%,均显著低于健康人【分别为(1.1±0.2)和(13.7±3.9)%,P<0.05】;21例肝组织G3/G4级患者TLR2 mRNA水平和细胞表面TLR2阳性的PBMCs百分比分别为(1.9±0.3)和(37.2±5.7)%,均显著高于26例G1/G2级患者【分别为(1.5±0.3)和(30.5±6.2)%,P<0.05】,而PBMCs CTLA-4 mRNA水平和细胞表面CTLA-4阳性的PBMCs百分比分别为(0.3±0.1)和(1.9±0.6)%,均显著低于G1/G2级患者【分别为(0.7±0.2)和(2.6±0.8)%,P<0.05】;17例肝组织F3/F4期患者TLR2 mRNA水平和细胞表面TLR2阳性的PBMCs百分比分别为(1.9±0.3)和(37.6±5.3)%,均显著高于25例F1/F2期患者【分别为(1.6±0.3)和(32.9±4.2)%,P<0.05】或5例F0期患者【分别为(1.3±0.2)和(28.7±1.9)%,P<0.05】,而PBMCs CTLA-4 mRNA水平和细胞表面CTLA-4阳性的PBMCs百分比分别为(0.3±0.1)和(1.8±0.6)%,均显著低于F1/F2期患者【分别为(0.6±0.2)和(2.3±0.7)%,P<0.05】或F0期患者【分别为(0.9±0.2)和(4.2±0.4)%,P<0.05】。结论 AIH患者PBMCs TLR2 mRNA和TLR2阳性的PBMCs百分比升高,而PBMCs CTLA-4 mRNA水平和CTLA-4阳性的PBMCs百分比降低,并与肝组织炎症活动度和纤维化程度相关,值得进一步研究。

Effects of Combination of 1,25(OH)_(2)D_(3) and TLR-4 Inhibitor on the Damage to HaCaT Cells Caused by UVB Irradiation

Objective Vitamin D and Toll-like receptor-4(TLR-4)inhibition are involved in the protection of keratinocytes.The effects of combination of 1,25(OH)_(2)D_(3) and TLR-4 inhibitor on the protection of keratinocytes against ultraviolet radiation B(UVB)irradiation remain unclear.This study was undertaken to explore the effects of combination of 1,25(OH)_(2)D_(3) and TAK-242(TLR-4 inhibitor)on the damage to HaCaT cells caused by UVB irradiation.Methods In vitro,HaCaT cells were treated with 1,25(OH)_(2)D_(3) or/and TAK-242 prior to UVB irradiation at the intensity of 20 mJ/cm^(2),then the production of reactive oxygen species(ROS),cell migration,apoptosis of cells,and the expression of oxidative stress,endoplasmic reticulum stress,and apoptosis related proteins were determined.Results Compared with the HaCaT cells treated with 1,25(OH)_(2)D_(3) or TAK-242,the cells treated with both 1,25(OH)_(2)D_(3) and TAK-242 showed,1)significantly lower production of ROS(P<0.05);2)significantly less apoptosis of HaCaT cells(P<0.05);3)significantly lower expression of NF-κB,Caspase-8,Cyto-C,Caspase-3(P<0.05).Conclusion The combination of 1,25(OH)_(2)D_(3) and TAK-242 could produce a better protection for HaCaT cells via inhibiting the oxidative stress,endoplasmic reticulum stress and apoptosis than 1,25(OH)_(2)D_(3) or TAK-242 alone.

Ureaplasma urealyticum-derived lipid-associated membrane proteins introduce IL-6, IL-8, and TNF-α cytokines into human amniotic epithelial cells via Toll-like receptor 2

Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on Toll-like receptors （TLRs）. Methods： LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells （HAECs） were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay （ELISA） and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results： LAMPs induced HAECs to produce inflammatory cytokines interleukin （IL）-6, IL-8, and tumor necrosis factor （TNF）-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor （anti-hTLR2-IgA）. Conclusions： LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.

类风湿性关节炎患者早期外周血单个核细胞Toll样受体2、4表达及意义

目的探讨类风湿性关节炎(RA)早期患者外周血单个核细胞(PBMC)Toll样受体(TLR)2、TLR4对其配体双糖链蛋白多糖(BGN)、脂多糖(LPS)的刺激反应性,阐明PBMC TLR2、TLR4在RA疾病早期中的作用。方法采用流式细胞术、实时荧光定量逆转录(RT)-聚合酶链反应(PCR)、酶联免疫吸附试验(ELISA)分别检测BGN和LPS刺激前后,RA组和健康对照组外周血CD14+单核细胞TLR4+细胞频率、PBMC TLR2 mRNA和TLR4mRNA及上清液白细胞介素6(IL-6)和肿瘤坏死因子α(TNFα)的含量变化。结果 RA早期患者TLR2 mRNA明显升高、TLR4 mRNA下降(P<0.01);经LPS刺激后,RA患者TLR4 mRNA升高3.50倍,而健康对照组下降到0.11倍;LPS和BGN促进了各组PBMC产生IL-6、TNFα,但RA早期患者组上升的倍数明显高于健康对照组。结论 PBMC TLR2、TLR4参与早期RA的发生、发展。

Toll样受体2mRNA在梅毒血清固定患者外周血单核细胞中的表达

目的研究梅毒血清固定患者外周血单核细胞(PBMCs)中Toll样受体2(TLR2)mRNA的表达情况,并探讨其与梅毒血清固定的发病关系。方法采用实时定量PCR方法检测TLR2 mRNA在观察组中40例梅毒血清固定患者PBMCs的表达水平,并以30例驱梅治疗后TRUST阴转者和30例正常人分别作为对照组。结果观察组PBMCs中TLR2 mRNA的表达水平(1.54±0.51)明显低于两对照组(2.49±0.86)和(2.45±0.74),差异有统计学意义(P均<0.05);两对照组PBMCs中TLR2mRNA的表达水平比较,差异无统计学意义(P>0.05)。结论梅毒血清固定患者PBMCs中TLR2mRNA的表达水平较驱梅治疗阴转者及正常人低,表明天然免疫异常可能导致梅毒血清固定的发生。

Lipopolysaccharide upregulates the expression of Toll-like receptor 4 in human vascular endothelial cells

OBJECTIVE: To investigate the expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA was extracted from ECV304 cells and isolated human umbilical vein endothelial cells (HUVECs) exposed to LPS, respectively. The quantification of TLR2 and TLR4 mRNA in HUVECs and EVC304 cells was carried out by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ECV304 cells and HUVECs were able to express TLR2 and TLR4 mRNA, but the expression levels of TLR4 appeared to be stronger than those of TLR2. LPS could upregulate the expression levels of TLR4 obviously, whereas it had no effect on the expression level of TLR2. CONCLUSIONS: Our data indicate that TLR4 may be the LPS signal transducer in endothelial cells and plays important roles in the cell activation of LPS. The ECV304 cell line is a better experimental model than isolated HUVECs in the research of endothelial cells.

商陆皂苷甲对银屑病患者外周血单个核细胞产生α肿瘤坏死因子和可溶性白介素2受体的影响

目的:研究在体外商陆皂苷甲(EsA)对银屑病患者外周血单个核细胞(PBMC)分泌肿瘤坏死因子(TNF-α)和可溶性白介素2受体(sIL-2R)水平的影响,探讨其治疗银屑病的可能机制。方法:标本取自30例寻常性银屑病患者和20名健康献血员,利用EsA分别与脂多糖(lipopolysaccharide,LPS)或植物血凝素(phytohemagglutinin,PHA)协同刺激PBMC,用双抗体夹心ABC-ELISA检测PBMC培养上清液中TNF-α和sIL-2R水平。结果:EsA浓度为1.0～10.0μg/mL时,呈剂量依赖性地明显抑制银屑病患者PBMC释放TNF-α(P<0.05)。EsA浓度在0.5～10.0μg/mL时,PBMC分泌sIL-2R呈轻微的下降趋势,但差异无统计学意义(P>0.05),EsA对PHA诱导的sIL-2R释放无显著的抑制作用。结论:抑制TNF-α等炎症递质的释放可能是中国商陆治疗银屑病的机制之一。

丙型病毒性肝炎患者PBMC mIL-2R的表达

目的 探讨丙型病毒性肝炎患者膜白介素 2受体 (mIL 2R)表达水平及其在丙肝转归中的作用。方法 用PCR和生物素 链霉亲和素法对 78例丙肝患者外周血单个核细胞 (PBMC)分别进行HCV RNA和植物血凝素诱导前后mIL 2R的检测。结果 丙肝患者PBMC静息期和诱导期mIL 2R表达水平分别为 (2 .94± 0 .88) %、(31.5 3± 3.38) % ,与正常对照相比 ,差异有显著性 (P <0 .0 1)。其中 ,急性丙肝患者静息期和诱导期mIL 2R表达水平分别为 (3.2 5± 0 .94 ) %、(32 .82± 3.84 ) % ,慢性丙肝患者静息期和诱导期mIL 2R表达水平分别为 (2 .77± 0 .84 ) %、(30 .97± 3.16 ) % ,两者相比差异有显著性 (P <0 .0 5 )。PBMC内HCV RNA(+)者静息期和诱导期mIL 2R表达水平分别为 (2 .37± 1.16 ) %、(30 .4 1± 4 .0 1) % ,PBMC内HCV RNA(- )者静息期和诱导期mIL 2R表达水平分别为 (3.2 1± 0 .80 ) %、(32 .15± 3.0 9) % ,两者相比差异有显著性 (P <0 .0 5 )。结论 丙肝患者体内mIL 2R水平降低 ,与丙肝的慢性化程度似有一定关系 ;HCV侵入PBMC后可进一步抑制mIL

动脉瘤性蛛网膜下腔出血患者外周血单个核细胞中瞬时受体电位通道M2水平与介入治疗预后的关系

目的 分析动脉瘤性蛛网膜下腔出血(aSAH)患者外周血单个核细胞中瞬时受体电位通道M2(TRPM2)水平与介入治疗预后的关系。方法 选取2019年8月至2021年5月在三亚市中医院接受介入治疗的148例aSAH患者作为研究对象。用Ficoll密度梯度离心法和蛋白质免疫印迹法检测外周血单个核细胞中TRPM2水平。用改良Rankin量表(mRS)评分评价介入治疗90 d后预后。用logistic回归分析TRPM2与aSAH患者介入治疗预后的关系。结果 根据随访后m RS评分将患者分为预后良好组(n=107)和预后不良组(n=41)。预后良好组年龄、Hunt-Hess分级Ⅲ级占比、脑血管痉挛占比和Glasgow昏迷量表(GCS)评分低于预后不良组(均P<0.05)。预后良好组治疗前后TRPM2水平低于预后不良组(均P<0.05)。治疗后TRPM2>0.28和Hunt-Hess分级Ⅲ级是a SAH患者预后的独立危险因素(P<0.05)。结论 aSAH患者介入治疗后TRPM2水平高与预后不良有关。

人Toll样受体2细胞外区基因重组腺病毒穿梭载体的构建及鉴定

目的:克隆人Toll样受体2(TLR2)细胞外区基因,并构建含有目的基因的重组腺病毒穿梭质粒pAdTrack-CMV-TLR2。方法:应用RT-PCR方法从人外周血单个核细胞中扩增TLR2细胞外区基因,克隆入pUCm-T载体并转化大肠埃希菌感受态DH5α,提取质粒进行酶切鉴定及DNA测序分析后,用KpnⅠ和HindⅢ双酶切,将目的片段定向克隆至经相同处理的pAdTrack-CMV腺病毒穿梭质粒中,双酶切鉴定并测序验证。结果:RT-PCR扩增得到约1000bp的目的基因片段,TA克隆后测序鉴定与GenBank中的序列一致,经酶切连接成功地将目的基因插入到腺病毒穿梭质粒pAdTrack-CMV中。结论:成功克隆了人TLR2细胞外区基因,并构建了含目的基因的重组腺病毒穿梭载体。

子宫内膜异位症患者外周血单个核细胞T细胞亚群、mIL-2R表达及临床意义

[目的]探讨子宫内膜异位症患者外周血单个核细胞(PBMC)T细胞亚群、mIL-2R表达及临床意义。[方法]用生物素-链霉亲素(BSA)法检测T细胞亚群和mIL-2R表达水平,以PHA体外诱导PBMC中mIL-2R表达。[结果]子宫内膜异位症患者CD3+、CD4+、CD8+及CD4+/CD8+比值分别为(38.9±7.0)%、(29.2±6.6)%、(23.9±4.2)%、1.3±0.5;PHA诱导前后mIL-2R表达水平分别为(3.9±1.2)%和(21.7±5.3)%,与正常对照相比,差异有统计学意义(P﹤0.5)。[结论]子宫内膜异位症患者存在定程度细胞免疫功能低下表现,T细胞活化障碍,并参与子宫内膜异位症的致病过程。

丙型肝炎患者外周血单核细胞中IFNAR2 mRNA表达的研究

目的探究丙型肝炎病毒(HCV)不同感染阶段,外周血单核细胞(PBMC)干扰素α/β受体2(IFNAR2)mRNA表达水平,并研究其与干扰素(IFN)疗效的关系。方法 58例丙肝抗体阳性患者,根据临床表现分为3组:急性肝炎组(4例),慢性肝炎组(46例),肝硬化组(8例)。15名健康志愿者作为对照组。应用密度梯度离心法将外周血单核细胞分离,应用RT PCR扩增IFNAR2 mRNA。结果 58例丙型肝炎患PBMC IFNAR2 mRNA检出率为87.9%,而对照组为33.3%,差异具有统计学意义(P<0.05);慢性丙型肝炎组较肝硬化组检出率高(93.5%和50%,P<0.05),2组间PBMC HCV RNA的阳性率无差别(P>0.05)。PBMC IFNAR2 mRNA水平与干扰素疗效相关(P<0.05)。结论丙型肝炎病毒感染可上调PBMC IFNAR2 mRNA的表达,慢性丙型肝炎患者PBMC IFNAR2 mRNA表达高于肝硬化患者,其表达可能与病毒载量无关,而与干扰素疗效相关。