Clinical significance of peripheral blood immune cells in patients with gastric cancer after surgery

BACKGROUND Gastric cancer is one of the most common malignant tumors worldwide,and surgical resection is one of the main ways to treat gastric cancer.However,the immune status of postoperative patients is crucial for prognosis and survival,and immune cells play an important role in this process.Therefore,it is helpful to understand the immune status of postoperative patients by evaluating the levels of peripheral blood immune cells,especially total T cells(CD3+),helper T cells(CD3+CD4+),and suppressor T cells(CD3+CD8+),and its relationship to sur-vival.AIM To analyzed the immune cells in peripheral blood of patients with gastric cancer after surgery,detect the levels of total T cells,helper T cells and suppressor T cells.METHODS A total of 58 patients with gastric cancer who received surgical treatment were included in the retrospective study.Flow cytometry was used to detect the level of peripheral blood immune cells and analyze the correlation between total T cells,helper T cells and inhibitory T cells.To explore the relationship between these immune markers and patient survival.RESULTS The results showed that the levels of total T cells,helper T cells,and suppressor T cells changed in patients after gastric cancer surgery.There was a significant positive correlation between total T cells,helper T cells and suppressor T cells(r=0.35,P<0.01;r=0.56,P<0.01).However,there was a negative correlation between helper T cells and suppressor T cells(r=-0.63,P<0.01).Follow-up showed that the survival rate of patients in the high-level total T cell group was significantly higher than that in the low-level group(28.87±24.98 months vs 18.42±16.21 months).The survival curve shows that the curve of patients in the high-level group is shifted to the upper right,and that of the low-level group is shifted downward.There was no significant difference between the levels of helper T cells and suppressor T cells and patient survival time.CONCLUSION By detecting peripheral blood immune cells with flow cytometry,we can initially evaluate the immune status of patients after gastric cancer surgery and initially explore its relationship with patient survival.

Flow cytometric detection of hepatitis C virus antigens in infected peripheral blood leukocytes: Binding and entry

AIM： We designed two synthetic-core-specific peptides core 1 （C1） and core 2 （C2）, and an E1-specific peptide （El）. We produced specific polyclonal antibodies against these peptides and used the antibodies for detection of HCV antigens on surface and within infected peripheral blood leukocytes. METHODS： Peripheral blood from a healthy individual who tested negative for HCV RNA was incubated with HCV type 4 infected serum for i h and 24 h at 37 ℃. Cells were stained by direct and indirect immunofluorescence and measured by flow cytometry. RESULTS： After 1 h of incubation, antibodies against C1, C2, and El detected HCV antigens on the surface of 27%, 26% and 73% of monocytes respectively, while 10%, 5% and 9% of lymphocytes were positive with anti-C1, anti-C2 and anti-E1 respectively. Only 1-3% of granulocytes showed positive staining with anti-C1, anti-C2 and anti E1 antibodies. After 24 h of incubation, we found no surface staining with anti-C1, anti-C2 or anti-E1. Direct immunostaining using anti-C2 could not detect intracellular HCV antigens, after 1 h of incubation with the virus, while after 24 h of incubation, 28% of infected cells showed positive staining. Only plus strand RNA was detectable intracellularly as early as 1 h after incubation, and remained detectable throughout 48 h post-infection. Interestingly, minus RNA strand could not be detected after 1 h, but became strongly detectable intracellularly after 24 h post-infection. CONCLUSION： Monocytes and lymphocytes are the preferred target cells for HCV infection in peripheral blood leukocytes. Our specific anti-core and anti-E1 antibodies are valuable reagents for demonstration of HCV cell cycle. Also, HCV is capable of infecting and replicating in peripheral blood mononuclear cells as confirmed by detection of minus strand HCV RNA as well as intracellular staining of core HCV antigen.

HLA Class I Expressions on Peripheral Blood Mononuclear Cells in Colorectal Cancer Patients

Objective： To investigate the expression change of human leukocyte antigen （HLA） class I on human peripheral blood mononuclear ceils （PBMCs） at both mRNA and protein levels, and to evaluate its roles in the development of colorectal cancer （CRC）. Methods： In the present study, 50 patients with CRC, 35 patients with benign colorectal lesion and 42 healthy volunteers were enrolled. Expression levels of HLA class I mRNA and protein were determined using real-time quantitative reverse transcription PCR （RT-PCR） and flow cytometry analysis, respectively. Results： The expression levels of HLA class I mRNA and proteins were not influenced by age and gender. The relative ratios of HLA class I mRNA were 0.99±0.27 in healthy controls, 0.76±0.19 in benign patients, and 0.48±0.21 in CRC patients. Mean fluorescence intensities of HLA class I were 145.58±38.14 in healthy controls, 102.05±35.98 in benign patients and 87.44±34.01 in CRC patients. HLA class I on PBMCs was significantly down-regulated at both mRNA and protein levels in patients with stage III and IV CRC. CRC patients with lymph node metastasis also showed a decreased HLA class I expression at protein level. Conclusion： HLA class I expressions on PBMCs are associated with staging of CRC and lymph node metastasis. Monitoring the expression of HLA class I on PBMCs may provide useful information for diagnosis and metastasis judgement of CRC.

EXPRESSION OF IL-2R ON PERIPHERAL BLOOD LYMPHOCYTES OF PATIENTS WITH COLORECTAL CANCER AND ITS CLINICAL SIGNIFICANCE

Objective: To study expression of membrane receptors of interleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%) in PBL of 105 colorectal cancer patients before operation and 100 normal individuals were examined by flow cytometer, and the results were clinically and pathologically analyzed. Results: The mean of CD25% in PBL of the normal individuals was 17.24±5.33, it was significantly lower (P<0.01) than that of the colon cancer patients (21.29±7.95) or rectal cancer patients (21.62±6.11). In contrast to the normal individuals, the means of CD25% in PBL in ulcer type (20.53±6.50) or protruded type (21.56±6.16) colorectal cancer patients were notably elevated (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was observed between the normal individuals and patients with less than 4 cm mass (22.10±5.43) or 4cm–8cm mass (20.90±6.96). The significant difference (P<0.05) of means of CD25% in PBL was also observed between the normal individuals and patients with greater than 8 cm mass (21.56±5.41). The mean of CD25% in PBL in patients with well differentiation colorectal cancer was 22.20±5.50, it was significantly higher than that in normal individuals (P<0.05). The means of CD25% in PBL in patients with middle or poor differentiation colorectal cancer were 21.30±6.89 and 22.15±5.71 respectively, they were obviously higher than that in normal individuals (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the colorectal cancer patients without metastatic lymph nodes (22.06±6.90) and normal individuals. The significant difference (P<0.05) of means of CD25% in PBL was present between the colorectal cancer patients with metastatic lymph nodes (20.73±6.40) and normal individuals. The means of CD25% in PBL in colorectal cancer patients in various clinic stages were significantly higher than that in the health subjects (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the patients whose ages were equal to or less than 60 (21.00±5.76) and normal individuals in the same age group. The significant difference (P<0.05) of means of CD25% in PBL was also present between the patients whose ages were greater than 60 (22.54±7.75) and normal individuals in the same age group. The significant difference (P<0.01) of means of CD25% in PBL was present between the male patients (22.55±7.05) and normal men. The significant difference (P<0.05) of means of CD25% in PBL was also present between the female patients (20.09±5.48) and normal women. Conclusion: The mean of CD25% in PBL of colorectal cancer patients was significantly higher than that in health subjects. Abnormally elevated CD25% were correlative with site of tumor growth, macropathology type of tumor, the degree of tumor differentiation, clinical stage and patient’s age and sex. It may be helpful to detect CD25% in PBL of colorectal cancer patients before operation for diagnosis, immune treatment and judging prognosis.

21色流式检测人非小细胞肺癌组织中免疫细胞亚群方案的建立

背景与目的肺癌组织的免疫微环境已成为关注的重点,随着多色流式的兴起,流式检测肺癌免疫微环境成为重要的手段,但多为检测细胞亚群占比或主要细胞亚群功能,无法同时对两者进行检测。因此本研究建立了一种可靠的21色流式方案,以检测人非小细胞肺癌(non-small cell lung cancer,NSCLC)肿瘤组织中免疫细胞各亚群。方法选用细胞膜表面抗体细胞分化簇(cluster of differentiation,CD)45、CD3、CD19、CD4、CD8、程序性死亡受体1(programmed cell death 1,PD-1)、CD39、CD103、CD25、CD127、趋化因子受体8(chemokine receptor8,CCR8)、CD56、CD11c、人类白细胞抗原(human leukocyte antigen,HLA)-DR、CD38、CD27、CD69、CD62L、CD45RA、CCR7和核酸染料L/D制定方案。首先对各抗体进行抗体滴定实验、电压优化、减一色染色和单色染色实验,确定各实验条件及检测方案后,采用6例健康成年志愿者外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)标本验证方案的可行性;检测分析6例NSCLC患者的肿瘤组织样本。结果采用建立的21色流式方案检测了6例NSCLC患者的肿瘤组织样本,可分析出肺癌组织中各细胞亚群的占比及主要细胞群的免疫表型和分化情况。结论成功建立的21色流式方案适用于PBMCs和NSCLC组织样本检测,为监测肺癌中免疫微环境状态提供了一种有效的新思路。

调节性T细胞在肝细胞癌患者外周血中的表达及临床意义

目的探讨在不同病理分期肝细胞癌(hepatocellular carcinoma,HCC)患者外周血中CD4^(+)CD25^(+)CD127^(LOW)Treg的含量变化及临床意义。方法收集60例HCC患者的外周抗凝血作为HCC试验组并进行Edmondson-steiner病理分期,20名健康体检者的外周抗凝血作为健康对照组,检测两组CD4^(+)CD25^(+)CD127^(LOW)Treg水平。结果HCC患者外周血中Treg占CD4^(+)T细胞的百分率显著高于健康对照组(P<0.01);且在HCC不同分期中,Ⅲ、Ⅳ期患者外周血中Treg占CD4^(+)T细胞的百分率显著高于Ⅰ、Ⅱ期患者(P<0.05)。结论在HCC患者外周血中,Treg随着疾病的发生发展而逐渐增多,且在Ⅲ、Ⅳ期快速增殖,其可能与肿瘤细胞免疫逃逸密切相关。

外周血涂片+骨髓涂片+流式细胞术联合应用对MDS早期诊断准确性的影响

目的 探讨在骨髓增生异常综合征(MDS)应用外周涂片联合骨髓涂片与流式细胞术进行早期诊断的临床效果。方法 选取2019年6月—2022年6月本院接收诊治的骨髓增生异常综合征患者120例作为此次研究对象,按照检验方式的不同将患者分为实验组和对照组,每组60例。其中,对照组单独使用外周血涂片进行MDS的早期诊断,实验组则联合外周血涂片、骨髓涂片和流式细胞术进行MDS的早期诊断。对比两组患者的诊断结果。结果 经过检查后,实验组患者的检测结果明显优于对照组,差异有统计学意义(P<0.05)。结论 将外周血涂片、骨髓涂片和流式细胞术联合起来进行MDS的早期诊断,检测准确性更高,能够为临床治疗提供更精准的数据支持。

The approaches in detecting cell cycle specificity of Fas-mediated apoptosis in leukemia cell lines and activated PBLs in vitro

Objective： To establish a system in detecting the cell cycle specificity induced by recombinant human Fas ligand in vitro, so as to provide a reliable platform for further exploring the mechanism of cell cycle control and regulation in Fas-mediated apoptosis. Methods： The target cells-leukaemia cell lines and activated peripheral blood lymphocytes stimulated by phytohemagglutinin were incubated with recombinant human Fas ligand for 6 to 36 h, apoptosis was detected by sub-G1, common annexin-Ⅴ/PI and modified annexin Ⅴ and propidium iodide （API） methods and analysed by flow cytometry. Results： The modified API method demonstrated that Fas-mediated apoptosis was cell cycle specific and initiated at G1 phase. The common annexinⅤ/PI method showed the most appropriate condition for the detection of typical cell cycle-specific apoptosis. The sub-G1 method could only illuminate late apoptosis and DNA histogram. Conclusion： Fas-mediated apoptosis was cell cycle-specific and initiated at G 1 phase. Based on the modified API and common AnnexinⅤ/PI methods, the establishment of stable and typical cell cycle-specific model in Fas-mediated apoptosis in vitro was feasible.

鸡外周血T淋巴细胞分群及免疫相关基因表达分析

为探索鸡抗病育种中简便实用的免疫功能测定指标,试验采用成像流式细胞术对四川山地乌骨鸡和大恒699肉鸡配套系的4周龄和10周龄外周血进行T淋巴细胞亚群分型,并检测外周血中白介素6(IL-6)、Toll样受体3(TLR3)和干扰素(IFN-γ)基因的表达水平。结果显示:10周龄时,CD_(4)^(+)T淋巴细胞亚群在两鸡种间差异显著(P<0.05),4周龄和10周龄的两鸡种CD_(8)^(+)T淋巴细胞亚群均有极显著差异(P<0.01);CD_(4)^(+)/CD_(8)^(+)比值在0.7~1.6之间呈正态分布,且随周龄增加而降低;大恒699肉鸡配套系的CD_(4)^(+)/CD_(8)^(+)比值比四川山地乌骨鸡高;IL-6和IFN-γ表达量随周龄增加而增加,但TLR3相反。细胞因子的增多和CD_(4)^(+)/CD_(8)^(+)比值的降低表明鸡只随周龄增长,机体免疫平衡发生改变。结果表明,基于成像流式细胞术进行鸡外周血T淋巴细胞分型简易可行,CD_(4)^(+)/CD_(8)^(+)比值可作为鸡免疫力检测的一个指标;天然免疫相关基因IL-6、TLR3和IFN-γ基因的表达量可以作为免疫力检测的参考。

Hot Spring Hydro Therapy Regulates Peripheral Leukocyte Together with Emotional Hormone and Receptor Positive Lymphocytes According to Each Constitution/Condition

Two primary defense systems develop in vertebrates, innate and adaptive. Despite those defense systems, the overwhelming problems of possessing this dual system, the innate or adoptive do not seem to guard or even prevent the development of one internal threat to survival, moreover sometimes direct to autoimmunity accompanying hypersensitivity. Further, every individual exposes to the risk of immunodeficiency in daily life with both internal and external stimuli. In this report, we tried to report the suitable menu for regulating the immune system modulate and discuss how we can compare and select each menu by evidence-based manner more than VAS. In a series of investigation, we have assessed the various health promoting menus such as hot spring hydrotherapy, acupuncture & moxibustion, light walking, independent to the western medicine. In this report, we try to summarize the regulatory effect of hot spring hydrotherapy as our representative menu under the influence of hypothalamus system. Their effects of elements were evaluated by the total number of peripheral leukocyte, its subsets, granulocyte and lymphocyte ratio, emotional hormones, receptor positive lymphoid cells. The regulative effects were confirmed also by other menu than hydrotherapy, acupuncture, walking etc. However, just by the simple comparison between the group before and after the hydrotherapy, we cannot exhibit the real regulatory effect by each menu. So we tried to show the effect by trying to indicate the constitution/condition dependent manner. Because the effect was not uniformly to each individual but it was dependent on each condition/constitution before the start of the menu. This is the one of the main proposes of this report for summaries that it is important to regulate the number and levels of each factor as ideal value. In other words, the regulation of each factor affected under the hypothalamus system is regulated with each constitution, under the influence of daily circumstances. The mode of regulation was the same in each menu, indicating that the higher leveler was down regulated and lower leveler was up regulated. Moreover, each vector of change was reversely correlated to the value of the day before. The other purpose of this study was designed to establish the regulatory effect of this hydrotherapy being not limited in the number of leukocyte in peripheral but in the emotional hormones. They were adrenaline and dopamine but not found another hormone for healthy individuals. Hot spring hydrotherapy for a short duration was expected to influence the immune system combining with hormone levels in the blood quantitatively. These hormones were evidenced by adrenalin and dopamine as well as adrenergic receptor positive lymphocytes. The final subject was to confirm and compare the effect of hot spring hydrotherapy and other menu. The reproducible effects were expressed by the linear slant and could compare each other with the value of slope. We tried to discuss the advantage and demerit, how we can compare each medicine with the peripheral leukocyte in number and function.

流式细胞术联合血红蛋白电泳法在孕妇胎母输血检测中的应用价值

目的:分析流式细胞术(FCM)联合血红蛋白电泳法(HBE)在孕妇胎母输血检测中的应用价值。方法:选取2020年5月—2022年5月解放军96605部队医院收治的60例疑似胎母输血综合征(FMH)患者作为研究对象。采集患者外周血,予以HBE、FCM、酸洗脱实验(KBT)检测,以KBT检测作为本研究结果的标准分析HBE联合FCM检测FMH的准确性与敏感性。结果:60例疑似FMH的孕妇中2例确诊,FCM联合HBE检测准确率为96.67%(58/60),敏感性为100.00%(2/2)。结论:FMH检测中使用FCM联合HBE检测具有较高的价值,对于胎儿出血量予以动态监测,能够为医务人员准确判断疾病、评估、严重程度及制定针对性治疗方案提供有效参考。

Laboratory Diagnosis of Acute Leukemia in Kenya: The Gaps and Opportunities

Acute leukemia (AL) is a malignant disease of the bone marrow in which hematopoietic precursors are arrested in an early stage of development. The diagnosis of leukemia and lymphomas, beyond morphology, is limited in low-resource countries including Kenya. Morphological diagnosis includes Cytological and Histological assessment of blood, bone marrow aspirates and tissues on suspected Acute leukemia patients. The World Health Organization (WHO, 2016) international guidelines on Acute leukemia diagnosis recommend that cytogenetic analysis, appropriate molecular genetics, Fluorescent in situ Hybridization (FISH) testing, and flow cytometric immuno-phenotyping should be done in addition to a morphologic assessment of Acute Leukemia. In facilities where resources are relatively available, immunophenotypic and genetic features have resulted not only in providing a more accurate leukemia diagnosis but also in identifying antigens or genes that can then be targeted for therapy. This article will look at the gaps in the diagnosis of Acute leukemia in low-resource settings like Kenya and opportunities available to improve diagnosis.

乳腺癌外周血微转移与远处转移的相关性

背景与目的:乳腺癌是一种全身性疾病,早期就可发生血行转移。本研究采用流式细胞仪定量检测乳腺疾病患者外周血肿瘤细胞含量,探讨外周血肿瘤细胞含量与远处脏器转移的相关性。方法:以人细胞角蛋白抗体和CD45抗体双色免疫标记,应用多参数流式细胞仪方法检测65例乳腺癌患者术前和10例乳腺良性病变及8例健康献血者外周血标本中肿瘤细胞含量。结果:65例乳腺癌患者中,57例检测出肿瘤细胞,阳性率为87.7%;乳腺良性病变、健康献血者外周血中未检测到肿瘤细胞。外周血肿瘤细胞阳性率与肿瘤T分期及N分期具有相关性(r=0.271,P=0.017;r=0.393,P=0.002)。除2例失访外,63例患者经过5年随诊,25例出现远处脏器转移,5年转移率为39.7%。所有远处转移病例均为外周血检测阳性患者,8例术前外周血检测阴性的患者均未发现远处转移,两组比较差异有统计学意义(P=0.014);而远处转移与肿瘤T分期、N分期均无明显相关性。结论:术前外周血肿瘤细胞数量与远处转移具有明显相关性,多参数流式细胞术检测乳腺癌患者外周血肿瘤细胞,对指导乳腺癌的个体化治疗具有潜在的应用价值。

外周血循环中肺癌细胞的流式细胞仪定量分析

目的 运用流式细胞仪定量分析外周血循环中肺癌细胞。方法 外周血经Ficoll梯度离心分离单核细胞组分,后者用CD45、细胞角蛋白(CK)及肺癌特异性单抗(2F7/S5A)染色后,应用流式细胞仪检测CD45- CK+ 2F7/S5A+ 细胞。然后计算每升血中上述细胞含量。结果 (1)外周血单核细胞的表型为CD45+ CK- 2F7/S5A- ,而小细胞和非小细胞肺癌细胞系均为CD45- CK+ 2F7/S5A+ 。因此,应用此方法能从106 白细胞中检测到1个肺癌细胞,敏感性达到每毫升血中检测到5 个肺癌细胞;(2)15 例正常人及7 例肺部良性疾患外周血中CD45- CK+ 2F7/S5A+ 细胞均为阴性,而40例肺癌患者外周血14 例发现阳性细胞,阳性率达35 % ,阳性细胞含量平均为1×105 /L。结论 本方法具有较好的敏感性和特异性。

肺癌患者外周血循环中癌细胞的流式细胞仪分析

目的 用流式细胞仪分析肺癌患者外周血循环中的癌细胞。方法 外周血经Ficoll梯度离心分离单核细胞组分 ,后者用CD45、细胞角蛋白 (CK)及肺癌特异性抗体 (2F7/S5A)染色后 ,应用流式细胞仪检测CD45 -CK+2F7/S5A+细胞。结果 检测了 16 5例肺癌患者外周血 ,发现 5 0例 (30 .30 % )患者外周血癌细胞呈阳性。其中非小细胞肺癌阳性率为 30 .6 7% (45 / 15 0 ) ,小细胞肺癌为 33 .33 %。阳性率与患者的病理分期有显著相关性 (P <0 .0 5 )。结论 利用流式细胞仪分析检测肺癌患者外周血循环中的癌细胞不仅有助于临床病理分期 ,而且对预测肺癌转移潜能具有应用价值。

不同冷冻保护剂对外周血干细胞保存的研究

目的:探索冻存外周血干细胞(peripheralbloodstemcell,PBSC)的最佳冷冻保护剂组合和最大细胞浓度。方法:将冷冻保护剂二甲基亚砜(dimethylsulfoxideDMSO)、羟乙基淀粉(hydroxyethylstarchHES)、人血清白蛋白(humanserumalbrinALb)及自体血浆(autoplasma)按不同比例组成3组冷冻保护剂,A组:5%DMSO+6%HES+4%ALb;B组:5%DMSO+6%HES+autoPLASMA;C组:5%DMSO+4%Alb。冻存PBSC的细胞浓度分别为1×108/ml、2×108/ml和4×108/ml3个梯度。比较3组不同冷冻保护剂对不同细胞浓度的PBSC的冻存效果。进行冻存前后粒单系集落形成单位集落形成能力(colonyformingunitGranulocytemacrophage和爆式红系集落形成单位CFUGM;burstformingUniterythroid,BFUE)、CD34+CD38-细胞的测定,及复苏后细胞拒染率测定。数据处理用统计学区组方差分析。结果:3组不同冷冻保护剂组合中以B组的CFUGM,BFUE回收率最高,在细胞浓度为1×108/ml的回收率分别为78.7±9.8%和69.8±14.1%;A组与C组的CFUGM回收率分别为68.3±6.2%和65.8±7.2%,BFUE回收率分别为63.4±9.7%和60.4±10.6%,B组与A组、C组方差分析F=56.7,0.01<P<0.05;A组与C组两组间无显著性差别。3组保护剂冻存PBSC复苏后的细胞拒染率及CD34+CD38-细胞的回收率分别在90%及80%以上。

动员外周血CD34阳性细胞流式细胞术测定中的策略探讨

探索用流式细胞术测定动员外周血中CD34^+细胞的简便有效的方法,以便在干细胞动员中进行有效的动态监测,及时收取干细胞,把握最佳移植效果。将经药物动员的移植供者的外周血用CD34和CD45荧光抗体标记,用红细胞裂解液将红细胞裂解后经流式细胞仪检测。运用恰当的分析窗口、有效的设门分析结合干细胞的生物学特性,确定CD34^+细胞在窗口中的确切位置,统计CD34^+细胞在有核细胞中的比例。结果表明,通过本方法可有效地测定0.05 ％-0.1％的CD34^+细胞的相对含量,在药物动员的第5或6天,多数患者外周血中CD34^+细胞达到峰值,某些患者外周血中CD34^+细胞可超过1％。结论:通过标记中的有效阻断和多参数分析,本方法可对动员外周血中微量的CD34^+细胞进行有效的动态监测,对适时采集干细胞进行移植提供了重要的参考。

肿瘤转移患者外周血细胞的某些生物学特征

目的:探讨肿瘤转移患者外周血细胞的生物学变化特征。方法:用流式细胞术对396例肿瘤转移患者和371例非转移患者外周血细胞的DNA倍体、凋亡水平和增殖活性进行了对比分析。结果:在有肿瘤转移的396例患者外周血细胞中,有45例出现了DNA异倍体,其检出率为11.36%;DNA异倍体患者血细胞的DNA指数(DNAindexDI)值为1.25±0.26。另371例非转移患者未发现DNA异倍体细胞。肿瘤转移者的细胞凋亡率和S期细胞比率均显著高于非转移者P<0.05,多器官转移者的细胞凋亡率和S期细胞比率也显著高于单器官转移者P<0.05。结论:肿瘤转移能引起患者外周血细胞中出现DNA异倍体,并使血细胞凋亡水平和增殖活性上调。

人外周血网织红细胞微核的流式细胞仪自动化检测

背景与目的 :建立一种基于单激光流式细胞仪的人外周血网织红细胞微核率自动化检测方法。材料与方法 :在本室已建立的大鼠骨髓嗜多染红细胞微核流式细胞仪自动化检测体系基础上 ,使用磁式细胞分选技术 ,富集并分离人外周血中的网织红细胞 ,采用吖啶橙(acridineorange,AO)荧光染色 ,用流式仪检测人外周血中的网织红细胞微核率。结果 :磁式细胞分选技术分离到的CD71阳性红细胞占网织红细胞总量的20 %至90 %,产量在不同个体之间存在显著差异 ,但同一个体其分离效果几乎没有差异 ;网织红细胞经过富集后的人外周血样本被分成明显的五个细胞群———网织红细胞、含微核的网织红细胞、成熟红细胞、含微核的成熟红细胞以及有核细胞 ;7名健康人外周血网织红细胞微核率平均值为2.36‰ ,10名接受累积放疗剂量0、4、10、20、30、40及50Gy后的鼻咽癌患者外周血网织红细胞微核率平均值分别为2.97‰、29.37‰、57.90‰、69.05‰、62.58‰、51.19‰及50.07‰。 结论 :成功建立了基于单激光流式细胞仪的人外周血网织红细胞微核率自动化检测体系。

流式单平台和双平台计数定量检测外周血淋巴细胞亚群的实验研究

目的建立并标准化流式细胞术外周血淋巴细胞亚群绝对含量的测定方法,探讨单平台和双平台法检测的特点与适用性。方法以三色流式细胞术,采用单平台和双平台法定量测定194例健康人外周血T细胞、B细胞、NK细胞亚群,并调查两种方法检测结果的精密性。结果单平台法健康人外周血总T细胞、T4细胞、T8细胞、T4/T8比值、DN-T细胞、DP-T细胞、B细胞和NK细胞的绝对含量依次为(1.69±1.10)、(1.10±0.74)、(0.73±0.60)、(1.51±0.62)、(0.15±0.10)、(0.02±0.03)、(0.36±0.32)和(0.37±0.41)×109/L;双平台法结果依次为(1.87±0.70)、(1.07±0.41)、(0.74±0.31)、(1.45±0.38)、(0.15±0.07)、(0.02±0.01)、(0.36±0.16)和(0.36±0.16)×109/L,两种方法检测结果之间差异无统计学意义(t检验,P>0.05)。双平台法检测结果的变异系数在4.5%～9.0%,均低于单平台法检测结果的变异系数值(F检验,P<0.01)。结论流式淋巴细胞亚群绝对计数的单平台法和双平台法均可用于临床检验,且单平台法检测结果的精密度更高,稳定性更好。