Metallothionein 1 Isoform Gene Expression Induced by Cadmium in Human Peripheral Blood Lymphocytes

Objective To study the gene expression of metallothionein 1 （MT-1） isoforms in human peripheral blood lymphocytes （HPBLs）. Methods The expression of mRNA representing the seven active MT-I genes was determined in HPBLs by quantitative RT-PCR before and after exposure to cadmium. Results Basal expressions of MT-IX, and MT-1A in HPBLs were similar to expression of housekeeping gene. In contrast, the basal gene expressions of MT- 1 H, IF, 1E, and 1G were a little transcripts in human HPBLs. No signal was detected for MT-1B. There was a sex difference （P〈0.05）. in basal gene expression of MT-1E. The levels of gene expression of MT-1A, 1E, IF, 1G, 1H, and 1X increased, but the level of MT-1B did not increase after exposure to cadmium. Conclusions Gene expressions of MT-1 G, MT-1 H, MT-1 F, and MT-1X in HPBLs can be used as a potential biomarker of cadmium exposure.

Expression of Heme Oxygenase-1 in the Peripheral Blood Mononuclear Cells from Asthmatic Patients

Summary： To explore the expression of heme oxygenase-1 （HO-1） in the peripheral blood mononuclear cells （PBMCs） and its relationship with pulmonary ventilation function in asthmatic patients, 18 asthmatic patients and 18 healthy subjects were selected. HO-1 protein and mRNA levels in PBMCs were measured by immunohistochemical staining and reverse transcription-polymerase chain reaction （RT-PCR）, respectively. Blood carbon monoxide Hb （COHb）, serum total IgE and pulmonary ventilatory function were observed. Our results showed that the percentage of cells positive for immunohistochemical staining of HO-1 were significantly higher in asthmatic patients （41.72±7.44） % than that in with healthy subjects （10.45±4.36）% （P〈0.001） and the optical density of PBMC HO-1 mRNA was higher in asthmatic patients （26.05±4. 14） than that in healthy subjects （10.82±4.26） （P〈0.001）. The relation analysis showed that PBMC HO-1 protein and mRNA levels had significantly negative relation with FEV, %, PEFR, MEFR50 , respectively （r=-0.51-0.89, P〈0.05-0. 001, respectively） and a positive relation with COHb and serum total IgE （r=0.48-0. 85, 0.05-0. 001, respectively）. It is concluded that the expression of PBMC HO-1 protein and mRNA increased significantly in asthmatic patients, and HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 may bear a relation with severity of asthma.

Expression of multidrug resistance 1 gene and C3435T genetic polymorphism in peripheral blood of patients with intractable epilepsy

BACKGROUND： Increased expression of multidrug resistance 1 （MDR1） mRNA in peripheral blood of patients with intractable epilepsy is not due to epilepsy drugs, but epilepsy behavior. Monitoring MDR1 expression in peripheral blood is a target for MDR1 gene evaluation. OBJECTIVE： To investigate the influence of antiepileptic drugs and seizures on MDR expression in intractable epilepsy, and to analyze the genetic polymorphisms of C3435T in the MDRl gene. DESIGN, TIME AND SETTING： Factorial designs and comparative observations at the experimental center of the Affiliated Hospital of Qingdao Medical College, Qingdao University between October 2003 and October 2004. PARTICIPANTS： A total of 120 subjects were recruited from the epilepsy clinical department of the Affiliated Hospital of Qingdao Medical College. Four groups （n = 30） were classified according to statistical factorial design： intractable epilepsy, treatment response, no treatment, and normal control groups. METHODS： One-step semi-quantitative reverse-transcription polymerase chain reaction technology was used to test expressions of the MDR1 gene in 120 subjects. C3435T polymorphisms in intractable epilepsy group and normal control groups were analyzed by polymerase chain reaction-restriction fragment length polymorphism. MAIN OUTCOME MEASURES： Expression of MDR1 mRNA in the four groups, and C3435T genetic polymorphisms in intractable epilepsy and normal control groups. RESULTS： MDRl gene expression was increased in the intractable epilepsy group, due to the factor seizures, but not the antiepileptic drugs. However, the interaction between the two factors was not statistically significant. Of the 30 subjects in the intractable epilepsy group, the following genotypes were exhibited： 3 （10%） C/C genotype, 9 （30%） C/T genotype, and 18 （60%） T/T genotype at the site of C3435T, while 4 （13%）, 10 （33%）, and 16 （53%） subjects were determined to express these genotypes in the normal control group, respectively. C and T allele frequency were 25% and 75% in the intractable epilepsy group, and 30% and 70% in the normal control group, respectively. However, there was no statistical difference between the groups. CONCLUSION： Results demonstrated that seizures, not antiepileptic drugs, induced MDR1 gene expression in intractable epilepsy. Genetic polymorphisms of C3435T in the MDR1 gene did not contribute to the development of multidrug resistance in patients with intractable epilepsy.

Changes of NLRP1-ASC-Caspase-1 signaling pathways in peripheral blood monocytes from patients with atrial fibrillation

Objective:To detect the expression of(Peripheral blood mononuclear cells,PBMCs)NLRP3 signal pathway in peripheral blood monocytes of patients with chronic heart failure,and to explore the expression of NLRP3 signal pathway and its induced inflammatory response in PBMCs of patients with different types of chronic heart failure.Methods:patients with chronic heart failure(NYHAⅡ~Ⅳ),Ⅱ(nude 20),Ⅲ(nude 20)andⅣ(nude 20)admitted to our hospital from 2019 to 2020 were selected,and 20 normal subjects were selected as the control group.The peripheral venous blood of all subjects was collected,and the plasma and monocytes were extracted respectively.The monocytes were identified by magnetic beads sorting.The mRNA and protein expression levels of NLRP3,ASC and Caspase-1 in PBMCs were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot,and the level of interleukin-1β(IL-1β)in plasma was detected by enzyme-linked immunosorbent assay((ELISA)).Results:compared with the normal control group,the expression of NLRP3,ASC,Caspase-1 protein and mRNA in PBMCs of patients with gradeⅡ,ⅢandⅣincreased.Compared with patients with gradeⅡ,the expression of these indexes increased in patients with gradeⅢandⅣ.Compared with patients with gradeⅢ,the expression of these indexes increased in patients with gradeⅣ.Compared with the normal control group,the plasma levels of IL-1βin patients with gradeⅡ,ⅢandⅣwere higher than those in patients with gradeⅡ,ⅢandⅣ(P<0.05).The expression of these indexes in patients with gradeⅢandⅣwas higher than that in patients with gradeⅢ(P<0.05).Conclusion:the results suggest that NLRP3-ASC-Caspase-1 signal pathway may cause chronic inflammation in patients with heart failure and play a role in the progression of chronic heart failure.

Expression of CK19 mRNA and MUC-1 mRNA in the peripheral blood of patients with colorectal cancer and their clinical significances

Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index.

Study on the relationship between the expression of NFκB1 and LncRNA-PACER in peripheral blood mononuclear cells of patients with pulmonary tuberculosis

Objective: To investigate the expression relationship between nuclear transcription factor kappa B1 (NFκB1) and long non-coding RNA PACER (LncRNA-PACER) in peripheral blood mononuclear cells (PBMCs) of patients with pulmonary tuberculosis. Methods: From February 2018 to March 2019, 40 patients with pulmonary tuberculosis (tuberculosis group) and 40 healthy persons (control group) were collected, the levels of TNF-α, IL-6 and IL-8 in serum were detected by enzyme-linked immunosorbent assay (ELISA);the expressions of LncRNA-PACER and NFκB1 mRNAs in PBMCs were detected by real-time fluorescence quantitative PCR;Western blot was used to detect the expressions of NFκB1 and COX 2 in PBMCs;Pearson method was used to analyze the expressions of LncRNA-PACER and NFκB1 in PBMCs of patients with pulmonary tuberculosis, and the expressions of LncRNA-PACER and NFκB1 in PBMCs of patients with pulmonary tuberculosis were analyzed. Results: Compared with the control group, the expressions of TNF-α, IL-6 and IL-8 in the serum of patients with pulmonary tuberculosis was significantly increased (P<0.05), and the expressions of LncRNA-PACER, NFκB1 mRNAs, proteins and COX-2 protein in PBMCs were significantly increased (P<0.05). The expressions of LncRNA-PACER and NFκB1 proteins in PBMCs were related to the number of pulmonary lesions and pulmonary cavity (P<0.05), and there was a positive correlation between the expression of LncRNA-PACER and the expression of NFκB1 mRNA in PBMCs of patients with pulmonary tuberculosis (r = 0.873, P<0.05). Conclusions: The expressions of NFκB1 and LncRNA-PACER in PBMCs of patients with pulmonary tuberculosis are significantly increased, they are positively correlated and both of them are related to the occurrence and development of pulmonary tuberculosis.

Lactobacilli,bifi dobacteria and E.coli nissle induce pro-and anti-inflammatory cytokines in peripheral blood mononuclear cells

AIM： To investigate whether the stimulation of peripheral blood mononuclear cells （PBMNC） with the cell debris and cell extraction of different probiotic strains is similar or species specific. METHODS： Three strains of bifidobacteria, 4 strains of lactobacilli, and E. coli nissle were sonicated and centrifuged in order to divide them into cell extract and cell debris. PBMNC were separated by density gradient and incubated for 36 h with either the cell debris or the cell extract of single strains of probiotic bacteria in doses from 10^2 to 10^8 CFU/mL. Cell supernatants were taken and interleukin （IL）-10, IL-1β, and tumor necosis factor （TNF）-α were determined by ELISA. RESULTS： Depending on the species super-family, the strains had different stimulation patterns. Except for both L. casei strains, the cell extract of bitTdobacteria and/actobacilli had less stimulating capacity than cell debris, whereas the cell extract of E. coli nissle had similar stimulating properties to that of the cell debris of the strain and significantly more stimulating capacity than that of bifidobacteria and lactobacilli. The cell debris of bifidobacteria stimulated more cytokine release than the cell debris of lactobacilli. The cell debris of lactobacilli did not have a stimulating capacity when lower concentrations were used. Neither cell extraction nor cell debris had an inhibitory effect on the production of the tested cytokines by stimulated PBMNC. CONCLUSION： The incubation of probiotic strains, which have been used in clinical trials for inflammatory diseases, with immunocompetent cells leads to different species specific reactions. High IL-10 response to cell debris of bifidobacteria and E. coli nissle can be found. This corresponds to positive effects of bihdobacteria and E. coli nissle in clinical trials for inflammatory bowel disease compared to negative outcomes obtained with lactobacilli.

Phosphorylation of Protein Kinase Akt by Mtorc2 in Peripheral Blood Mononuclear Cells of Patients with Cancer and Diabetes

Akt/mTOR/p70S6K1 signaling pathway plays an important role in the pathogenesis of cancer and diabetes.Macrophages and lymphocytes are involved in the pathogenesis of diabetes,diabetic atherosclerosis,formation of insulin resistance as well as immune response to cancer and tumor maintenance.The aim of the study was to determine the Akt activation by mTORC2 in peripheral blood mononuclear cell(PBMC)of patients with type 2 diabetes and cancer.The following groups were studied:control group,patients with type 2 diabetes,cancer patients and patients with both cancer and diabetes.The amounts of phospho-Akt(р-S473)and phospho-p70S6K1(p-T389)were determined using ELISA kits.The amount of phosphorylated Akt significantly increases in PBMC of patients with cancer.There was no effect in PBMC from patients with type 2 diabetes and significant decrease in the amount of phospho-Akt in PBMC of the patients group both with cancer and diabetes.p70S6K1 activation was observed in PBMC of the groups 2 and 3 patients.Thus,chronic diseases such as type 2 diabetes and cancer can affect the signaling mechanisms in blood cells.The state of Akt phosphorylation in leukocytes can indicate the activity of mTORC1 and its substrates,which may be important for the evaluation of the pathological process and the efficacy of the drugs.

脓毒症患者外周血单核细胞PD-L1/PD-1表达及临床意义

目的观察脓毒症患者外周血单核细胞(PBMC)的程序性细胞死亡因子-1(programmed cell death-1,PD-1)及其配体(programmed cell death-1 ligand,PD-L1)表达变化并探讨其临床意义。方法收集2015-01~2015-10入住我科的脓毒症患者38例,对照组为健康人15例。采集脓毒症患者外周血,采集时间点为入住ICU第1天、第3-4天、第7-8天,分离PBMC。流式细胞技术检测单核细胞、树突细胞PD-L1表达及CD8^+T淋巴细胞PD-1表达。并记录相应时间点中性粒细胞和淋巴细胞绝对计数。结果与对照组相比,脓毒症患者入住ICU第1天、第3-4天及第7-8天的单核细胞PD-L1表达和CD8^+T淋巴细胞均明显升高(P<0.01);脓毒症患者入住ICU第1天、第3-4天树突细胞PD-L1表达均明显升高(P<0.05)、第7-8天的树突细胞PD-L1表达有升高趋势,但差异没有统计学意义(P>0.05)。Spearman相关分析,树突细胞PDL1表达与中性粒细胞绝对计数呈正相关(r_s=0.272,P=0.018)。结论患者PBMC的PD-L1/PD-1上调表达可能反映脓毒症诱导的免疫抑制。

可溶性PD-L1及CTLA-4在原发性肝癌伴恶性腹腔积液患者中的表达水平及临床意义

目的研究原发性肝癌(PLC)伴恶性腹腔积液(MA)患者腹腔积液、外周血及单个核细胞(PBMC)中可溶性程序性死亡配体-1(sPD-L1)、可溶性细胞毒T淋巴细胞相关抗原-4(sCTLA-4)的表达水平,初步探讨其临床意义。方法选择2016年5月至2019年5月首都医科大学附属北京佑安医院肝病免疫科收治的189例腹腔积液患者作为研究对象,其中92例伴有MA的PLC患者为A组,97例伴有腹腔积液的肝硬化患者为B组。采用酶联免疫吸附法(ELISA)检测患者腹腔积液及外周血sPD-L1、sCTLA-4的表达水平,并采用聚合酶链反应(PCR)方法检测患者外周血PBMC中PD-L1、CTLA-4 mRNA的表达水平。分析sPD-L1、sCTLA-4联合检测对PLC伴MA的诊断价值。结果 A组患者PBMC中PD-L1、CTLA-4 m RNA表达水平显著高于B组,外周血及腹腔积液中sPD-L1及sCTLA-4表达水平显著高于B组(P<0.05);外周血sPD-L1、sCTLA-4表达水平与外周血PBMC中PD-L1、CTLA-4 mRNA表达水平呈显著正相关(P<0.05);腹腔积液与外周血中sPD-L1、sCTLA-4表达水平呈显著正相关(P<0.05);腹腔积液中sPD-L1与sCTLA-4联合检测诊断PLC伴MA的灵敏度、特异度分别为91.4%、95.2%。结论 PLC患者腹腔积液、外周血及外周血PBMC中PD-L1与CTLA-4的表达与病因密切相关,可以在一定程度上反映不同原因腹腔积液导致的免疫微环境差异,有望成为PLC伴MA诊断及免疫治疗的靶点。

与猪PD-1和PD-L1互作表位多肽的筛选及其免疫调节功能

本试验旨在筛选和鉴定与猪程序性死亡因子1(PD-1)及其配体(PD-L1)相互作用的表位多肽,为阻断猪PD-1/PD-Ls通路逆转机体的免疫功能提供新策略。根据已解析人与鼠的PD-1与PD-L1相互作用的关键氨基酸位点信息,分析猪PD-1与PD-L1蛋白相互结合的关键氨基酸位点,在关键氨基酸位点处设计系列表位多肽,固相合成法合成表位多肽;分离自然感染猪圆环病毒2型(PCV2)仔猪的外周血单个核细胞(PBMC),检测表位多肽与猪重组蛋白PD-1、PD-L1和PBMC的结合能力,选取能结合猪PD-L1和PBMC的表位多肽pPD-15,检测其对猪PBMC增殖的影响,分析其作为佐剂免疫后对小鼠猪瘟病毒(CSFV)抗体水平的影响,最后,高效液质联用色谱法(HPLC-MS)检测表位多肽pPD-15的纯度和氨基酸序列正确性。结果显示,表位多肽pPD-15能结合猪PD-L1和PBMC;增殖试验显示,pPD-15组M1的平均荧光强度(74.20%)比空白对照组M1的平均荧光强度(4.37%)提高了69.83%,表明表位多肽pPD-15可促进猪PBMC的增殖;动物免疫试验显示,pPD-15作为佐剂可以提高CSFV抗体水平;HPLC-MS结果显示,合成的表位多肽pPD-15纯度高,氨基酸序列正确。本试验证实,表位多肽pPD-15具有提高体内外免疫力的能力,为研制新型免疫调节佐剂提供了科学依据。

金属肽酶含血小板反应蛋白1作为妊娠剧吐诊断标志物的可行性研究

目的研究金属肽酶含血小板反应蛋白1(ADAMTS-1)作为妊娠剧吐(HG)诊断标志物的可行性。方法选取海口市人民医院2019年1月至2022年1月收治的100名孕妇作为研究对象。诊断为妊娠剧吐的49例患者作为实验组,另外51名健康孕妇为对照组。比较两组外周血清的ADAMTS-1表达水平及孕妇的一般资料、临床特征和生化指标,并评估外周血清ADAMTS-1表达水平和尿酮症之间的相关性。结果实验组的尿酮、血糖、谷丙转氨酶、中性粒细胞计数、血小板比积、血小板平均体积、降钙素原、血小板分布宽度和ADAMTS-1水平高于对照组且具有显著统计学意义(t值分别为0.000、-2.490、-2.780、-3.024、-2.674、-2.978、-7.595、-19.812、-11.596,P<0.05);ADAMTS-1与患者尿酮、谷丙转氨酶、中性粒细胞计数、血小板平均体积、降钙素原、血小板分布宽度显著正相关(r值分别为0.58、0.22、0.21、0.22、0.51、0.66,P<0.05),与患者红细胞平均体积和促甲状腺激素显著负相关(r值分别为-0.23、-0.32,P<0.05);使用ADAMTS-1诊断妊娠剧吐的受试者工作特征曲线(ROC)曲线下面积(AUC)为0.94(95%CI=0.88~1.00,P<0.05),ADAMTS-1的临界值为9.48ng/mL,灵敏度为85.71%,特异性为98.04%。结论HG患者的血清ADAMTS-1显著升高,可作为妊娠剧吐的有效生物标志物。

猪圆环病毒2型对外周血单核细胞中PD-1,PD-L1和IL-21转录水平的影响

用猪圆环病毒2型(PCV2)体外感染外周血单核细胞,建立荧光定量PCR方法检测PCV2的病毒载量;同时,在PCV2感染后12 h、24 h、36 h、48 h、60 h、72h收集细胞,抽提RNA进行反转录,检测PD-1、PD-L1和IL-21的转录水平变化,分析评价PCV2感染对PD-1、PD-L1和IL-21的转录水平变化的影响。结果显示,PD-1在感染外周血单核细胞后72 h显著升高,达到峰值;PD-L1在感染后转录水平都显著升高,48 h达到峰值;IL-21在感染后48 h转录水平最低。结果表明,PCV2不仅能够在体外感染外周血单核细胞,而且随着病毒载量的增加,导致PD-1、PD-L1和IL-21的转录水平显著升高。以上结果表明,PCV2感染导致PD-1、PD-L1和IL-21的转录水平升高,通过激活PD-1/PD-L1通路,从而抑制IL-21的转录水平;相反,IL-21刺激PD-1、PD-L1的转录水平升高。研究结果为探索PCV2的致病机制和控制PCV2的感染提供了理论基础。

白细胞介素1受体颉颃剂抑制脂多糖促奶牛外周血单个核细胞氧化应激损伤作用的研究

试验以脂多糖(LPS)为刺激源,以细胞活力、抗氧化指标和炎症因子为判断指标,探讨白细胞介素1受体颉颃剂(IL-1Ra)通过抑制白细胞介素1β(IL-1β)的活性,对LPS诱导外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs)氧化损伤的缓解作用。试验采用单因子完全随机设计,PBMCs被随机分为7个组(每组6个重复),分别给予不同的处理:第1组是阴性对照组(Neg组),完全培养基培养30 h;第2组损伤组(Dam组),是在完全培养基中培养6 h后,再经10μg/mL的LPS工作液培养24 h;第3至7组(R0.25、R0.5、R1、R5组和R10组)细胞分别经浓度为0.25、0.5、1、5、10 ng/mL的IL-1Ra培养6 h,接着经10μg/mL的LPS工作液培养24 h。结果表明:与Neg组相比,Dam组的细胞活力、抗氧化相关酶[包括总抗氧化能力(T-AOC)以及总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和硫氧还蛋白还原酶(TrxR)]的活性显著降低,丙二醛(MDA)浓度、炎症因子白细胞介素-6(IL-6)和IL-1β含量以及诱导型一氧化氮合酶(iNOS)活性、一氧化氮(NO)含量均显著升高(P≤0.05)。与Dam组相比,R1组显著逆转了氧化损伤引起的上述抗氧化活性的降低和炎症因子浓度的升高,其他IL-1Ra处理组对上述指标的逆转效果不同程度地低于R1组(P≤0.05)。上述结果说明,LPS通过诱发PBMCs产生大量IL-1β进而导致细胞氧化损伤,IL-1Ra剂量依赖性地缓解了LPS引起的氧化损伤,添加剂量以1 ng/mL为宜。

PD-1、PDL1在宫颈癌与宫颈病变外周血中的表达水平及意义分析

目的观察程序性死亡受体1(PD-1)与其配体1(PD-L1)在宫颈癌或宫颈病变患者外周血的表达水平并分析其临床意义。方法回顾性选取2015年1月到2017年1月收治的85例宫颈癌患者为宫颈癌组,40例宫颈上皮内瘤样病变(CIN)患者为CIN组,30例健康者为对照组。采集外周静脉血,用流式细胞仪检测外周血样本中PD-1^+CD4^+、PD-1^+CD8^+T细胞和PD-L1^+CD14^+单核细胞的比率,分析其表达与宫颈癌患者临床病理特征及预后的关系。利用ROC曲线分析外周血PD-1^+CD4^+T细胞、PD-1^+CD8^+T细胞、PD-L1^+CD14^+单核细胞对宫颈癌、CIN的鉴别效能。对影响宫颈癌患者预后的因素进行单因素和多因素分析。结果①宫颈癌组患者外周血PD-1^+CD4^+T细胞、PD-1^+CD8^+T细胞比率均高于CIN、对照组,PD-L1^+CD14^+单核细胞比率高于对照组,差异有统计学意义(P<0.05)。②临床分期Ⅲ期、低分化、肿瘤≥4 cm、有淋巴结转移、有宫旁浸润的宫颈癌组患者外周血PD-1^+CD4^+T细胞、PD-1^+CD8^+T细胞表达水平明显高于Ⅰ期或Ⅱ期、中高分化、肿瘤<4 cm、无淋巴结转移、无宫旁浸润者,差异有统计学意义(P<0.05)。③ROC曲线分析外周血PD-1^+CD4^+T细胞、PD-1^+CD8^+T细胞、PD-L1^+CD14^+单核细胞对宫颈癌、CIN的鉴别效能,曲线下面积(AUC)分别为0.734(95%CI:0.643~0.825)、0.791(95%CI:0.705~0.878)、0.597(95%CI:0.492~0.703)。④淋巴结转移(OR=1.684)、PD-1^+CD4^+T细胞高表达(OR=1.379)、PD-1^+CD8^+T细胞高表达(OR=1.499)是影响3年无进展生存率独立因素(P<0.05)。结论PD-1、PD-L1在宫颈癌、CIN外周血中上调表达,并与宫颈癌患者临床病理特征有关,且PD-1^+CD4^+T细胞、PD-1^+CD8^+T细胞能较好鉴别宫颈癌、CIN。

HBV相关性肝病患者外周血PBMCs表面Tim-3、PD-1表达水平检测及意义

目的分析乙型肝炎病毒(HBV)感染后相关肝病患者外周血单个核细胞(PBMCs)表面T细胞免疫球蛋白黏蛋白分子-3(Tim-3)、程序性死亡受体-1(PD-1)表达水平变化及意义。方法选取2017年1~12月川北医学院附属医院HBV携带者20例,慢性乙型肝炎(CHB)患者30例,重型乙型肝炎患者20例,乙肝肝硬化患者30例,肝细胞癌患者20例,以同期20例健康体检者作为健康对照组。检测各研究对象外周血PBMCs表面Tim-3、PD-1表达水平。结果 PBMCs表面Tim-3、PD-1表达水平在健康对照组最低,与HBV携带组比较差异无统计学意义(P>0.05),随着病情加重,PBMCs表面Tim-3、PD-1表达水平逐渐升高,在重型乙型肝炎组、肝细胞癌组最高,各组患者PBMCs表面Tim-3、PD-1表达水平与健康对照组、HBV携带者组比较,差异有统计学意义(P<0.05);HBV感染患者PBMCs表面Tim-3、PD-1表达水平与HBV DNA载量呈负相关(r=-0.431和-0.422,均P<0.05),与ALT、AST水平呈正相关(r=0.214、0.325、0.234和0.354,均P <0.05);HBV感染患者总体PBMCs表面Tim-3的表达水平与PD-1表达量呈正相关(r=0.967,P <0.05)。结论免疫负性调节因子Tim-3、PD-1与HBV相关性肝病患者肝组织炎症及纤维化发生相关,对Tim-3、PD-1水平调节可能为其临床免疫治疗提供新思路。

乙型肝炎患者恩替卡韦抗病毒治疗后外周血T淋巴细胞PD-1的表达及其与HBeAg血清学的关系

目的:探讨恩替卡韦对慢性乙肝(CHB)患者外周血T淋巴细胞表面程序性死亡受体1(PD-1)表达水平的影响及其与CHB患者血清学指标的关系。方法:选取2012年1月~2013年1月在本院接受恩替卡康治疗的35例血清HBeAg阳性的CHB患者为研究对象,分别于恩替卡韦治疗前、治疗1个月、3个月、6个月、12个月抽取静脉血5mL,采用荧光定量PCR检测患者血清HBV DNA载量,流式细胞术检测T淋巴细胞PD-1表达水平,全自动化生化分析仪测定患者血清谷草转氨酶(ALT)水平。结果:35例CHB患者经恩替卡韦治疗后HBeAg转换18例,HBeAg未转换17例。与HBeAg未转换组相比,HBeAg转换组治疗1个月、3个月、6个月、12个月HBV DNA载量、ALT水平、CD4^+T细胞PD-1、CD8^+T细胞PD-1水平较低,差异有统计学意义(P<0.05)。两组患者治疗前CD4^+T细胞表面PD-1、CD8^+T细胞表面PD-1与HBV DNA载量、ALT水平无相关性(均P>0.05),而治疗后CD4^+、CD8^+T细胞表面PD-1与HBV DNA载量、ALT水平呈正相关(P<0.05)。结论:恩替卡韦能有效抑制CHB患者HBV DNA复制,改善患者肝功能。外周血CD4^+、CD8^+T细胞表面PD-1可作为CHB患者经恩替卡韦治疗后血清学转换的预测指标。

PD-1和PD-L1在透明细胞肾细胞癌患者外周血和肿瘤组织中的表达及相关性

目的探讨透明细胞肾细胞癌(cc-RCC)患者外周血和肿瘤组织中程序性细胞死亡受体1(PD-1)/程序性细胞死亡配体1(PD-L1)的表达水平及其相关性。方法采用双抗体夹心ABC-ELISA法检测48例cc-RCC患者与10例正常外周血sPD-1和sPD-L1,采用免疫组化检测48例cc-RCC肿瘤组织和10例正常肾组织中PD-1和PDL1的表达。结果cc-RCC组外周血sPD-L1均数明显高于对照组(4.17 ng/ml对0.86 ng/ml,P<0.01),PD-1/PDL1在cc-RCC肿瘤组织的阳性率明显高于对照正常组织的阳性率(39.6%/45.8%对0,P<0.01),肿瘤组织中PDL1表达与外周血sPD-L1水平呈正相关性(相关系数r=0.721,P=0.000)。结论PD-1和PD-L1参与cc-RCC的病理过程,肿瘤组织中PD-L1高表达可能会使外周血sPD-L1水平升高。

食管鳞癌外周血细胞PD-L1、PD-1及TCF-1 mRNA表达与预后的关系

目的探讨外周血细胞中程序性死亡配体-1(PD-L1)、程序性死亡受体(PD-1)及TCF-1 mRNA变化及其在食管鳞癌(ESCC)诊断和预后中的价值。方法回顾性分析2019年1月至2019年12月间河南科技大学第一附属医院临床医学院肿瘤医院病理确诊的91例ESCC患者和63例非癌对照外周血细胞中PD-L1、PD-1和TCF-1的mRNA表达变化,并通过ROC曲线分析诊断ESCC效能。Log-rank检验确定不同组别生存曲线差异,确定PD-L1、PD-1和TCF-1 mRNA表达与总生存期相关性。结果ESCC患者外周血细胞中PD-L1、PD-1和TCF-1 mRNA显著高于非癌对照者(P<0.05),其诊断ESCC的敏感性和特异性分别为0.58/0.68、0.92/0.64和0.95/0.58,AUC分别为0.66、0.84和0.78,且这3个mRNA分子表达呈正相关。PD-L1、PD-1、TCF-1 mRNA高表达的ESCC患者总生存期明显高于低表达者(P<0.05)。结论PD-L1、PD-1和TCF-1是ESCC潜在的诊断和预后生物标志物分子。

PD⁃1抑制剂联合全身化疗治疗复发转移性宫颈癌的临床效果观察

目的探讨程序性细胞死亡受体-1(PD-1)抑制剂(卡瑞利珠单抗)免疫治疗联合全身化疗治疗复发转移性宫颈癌的临床效果。方法选择2019年2月—2021年6月定州市人民医院收治的复发转移性宫颈癌64例,依据随机数字表法分为研究组和对照组各32例。对照组给予紫杉醇联合顺铂全身化疗方案,研究组给予全身化疗方案+PD-1卡瑞利珠单抗免疫治疗。比较2组临床疗效,分析治疗前及治疗3个周期后鳞状细胞癌抗原(SCC)、外周血淋巴细胞/单核细胞(LMR)及血小板/淋巴细胞(PLR)指标水平及Kamofsky评分变化,并观察治疗期间毒性作用发生情况及随访期间患者总生存期。结果研究组总有效率、疾病控制率分别为93.75%(30/32)、96.88%(31/32),高于对照组的68.75%(22/32)、75.00%(24/32),差异有统计学意义(P<0.05)。治疗3个周期后,2组血清SCC、PLR水平较治疗前降低,LMR较治疗前升高,且研究组改善程度优于对照组(P<0.05)。治疗后,2组Kamofsky评分均较治疗前升高,且研究组高于对照组(P<0.05)。治疗后研究组1、2年生存率及总生存期高于或长于对照组(P<0.05)。2组毒性作用多数为1~2级。研究组血小板下降和转氨酶升高比例分别为37.50%(12/32)和28.12%(9/32),高于对照组的18.75%(6/32)和9.38%(3/32),差异有统计学意义(P<0.05);2组贫血、白细胞下降、恶心、腹泻、乏力等毒性作用发生率比较差异无统计学意义(P>0.05);研究组中发生反应性毛细血管增生症、甲状腺功能减退、皮疹、带状疱疹、过敏等经对症处理后症状消失。结论PD-1抑制剂联合全身化疗治疗复发转移性宫颈癌提高了临床效果、生存质量及生存率,延长生存期,改善了机体的炎症免疫反应状态,毒性作用较少,患者耐受性好。