L_(9)(3~4)正交试验法优选肺炎合剂制备工艺

目的 优选肺炎合剂的制备工艺。方法 以盐酸麻黄碱含量及干膏率的综合评分为评价指标,以加水量、煎煮时间、煎煮次数为考察因素,采用L_(9)(3~4)正交试验法优选肺炎合剂水提取工艺,采用单因素试验法考察其浓缩温度及纯化工艺,并进行验证试验。结果 优选最佳制备工艺为煎煮2次,第1次加8倍量水,第2次加6倍量水,每次煎煮1.5 h,煎液减压(压强为0.05 mbar)浓缩温度控制在55~85℃,采用离心机(转速为10 000 r/min,离心半径为92 mm)过滤。按优选工艺制备的3份样品中,盐酸麻黄碱含量为0.14 mg/m L,干膏率为10.66%。结论 优选工艺科学、合理,操作简便,且稳定性、重复性好,可为肺炎合剂的工业化生产提供试验依据。

L_(9)(3^(4))正交试验法优选3种静脉用难溶性抗菌药物调配方法

目的优选哌拉西林钠他唑巴坦钠、头孢呋辛钠、头孢哌酮钠舒巴坦钠3种静脉用难溶性抗菌药物的调配方法。方法以溶剂品种、初溶溶剂体积、振荡时间为影响因素,以调配液中不溶性微粒数量为考察指标,采用L_(9)(3^(4))正交试验法优选3种静脉用难溶性抗菌药物的调配方法。结果溶剂种类(灭菌注射用水、0.9%氯化钠注射液、5%葡萄糖注射液)对3种药品不溶性微粒数量均无显著影响(P<0.05)。哌拉西林钠他唑巴坦钠的优选调配方法为采用22.5 mL溶剂预溶后,置振荡器上振荡120 s,稀释,调配;头孢呋辛钠的优选调配方法为采用12.5 mL溶剂预溶后,置振荡器上振荡40 s,稀释,调配;头孢哌酮钠舒巴坦钠的优选调配方法为采用10.0 mL溶剂预溶后,置振荡器上振荡40 s,稀释,调配。以0.9%氯化钠注射液为溶剂,按优选方法分别调配哌拉西林钠他唑巴坦钠、头孢呋辛钠、头孢哌酮钠舒巴坦钠,3种药品的不溶性微粒数量均显著减少(P>0.05)。结论优选的调配方法稳定、可行,可保障静脉输液质量。

基于信息熵赋权法的L_(9)(3^(4))正交试验优选治痤方水提工艺

目的优选治痤方的水提工艺。方法以提取次数、加水量、提取时间为考察因素,通过信息熵赋权法计算黄芩苷、丹酚酸B的含量和浸膏得率的权重系数,采用L_(9)(3^(4))正交试验优选治痤方的水提工艺。结果黄芩苷、丹酚酸B的含量和浸膏得率的信息熵分别为0.9751,0.9802,0.9810,权重系数分别为0.3909,0.3108,0.2983。最佳提取工艺为饮片加10倍量水,提取3次,每次1.5 h。验证试验结果显示,黄芩苷和丹酚酸B的平均含量分别为3.32,3.24 mg/mL,平均浸膏得率为28.42%,平均综合评分为0.9945。结论所优选的治痤方水提工艺稳定、可靠,可为治痤方的提取工艺和质量研究提供参考。

L_(9)(3^(4))正交试验法优选黄芩解毒颗粒水提取工艺

目的优选黄芩解毒颗粒的水提取工艺。方法以加水量、煎煮时间和煎煮次数为考察因素,以干膏得率和主药黄芩中指标性成分黄芩苷的回收率为评价指标,采用L_(9)(3^(4))正交试验优选提取工艺,并进行验证试验。结果最佳提取工艺为加8倍量水,煎煮2次,每次1.0 h。所制备样品的平均干膏得率和黄芩苷回收率分别为27.51%和44.65%,RSD分别为0.93%和0.61%(n=3)。结论优选的提取工艺操作简便、工艺稳定,可用于黄芩解毒颗粒的工业化生产。

Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

外周血CD3^(+)CD4^(-)CD8^(+)T细胞与T_(1)期高级别尿路上皮癌预后的关系

目的研究外周血CD3^(+)CD4^(-)CD8^(+)T细胞数量与T_(1)期高级别尿路上皮癌术后预后的相关性。方法回顾性分析2012年1月至2016年12月在上海市第一人民医院诊治为T_(1)期高级别尿路上皮癌的患者168例,以外周血细胞数量为320个/μl分界将患者分为CD3^(+)CD4^(-)CD8^(+)正常组(87例)和数量低组(81例)。采用Kaplan-Meier法制作生存曲线,Cox回归模型分析外周血CD3^(+)CD4^(-)CD8^(+)T细胞数量与T_(1)期尿路上皮癌的无复发生存和无进展生存(PFS)的关系。结果CD3^(+)CD4^(-)CD8^(+)正常组和CD3^(+)CD4^(-)CD8^(+)数量低组的PFS均值分别为54.97个月vs 49.07个月,外周血CD3^(+)CD4^(-)CD8^(+)T细胞数量低和更短的PFS相关(P=0.017,HR=2.31,95%CI=1.163~4.570)。结论外周血CD3^(+)CD4^(-)CD8^(+)T细胞数量低是T_(1)期尿路上皮癌进展的独立危险因素。

Effect of macrophage polarization regulated by miR-29b,B7H3 on CD4^(+)T cell differentiation in asthma

Objective:To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4^(+)T.Methods:1.PBMC was extracted from peripheral blood mononuclear cells of children with asthma and normal children in the affiliated Children's Hospital of Soochow University,and RNA was extracted and reverse transcribed.The expression of miR-29b and B7H3mRNA was determined by real-time quantitative polymerase chain reaction(Q-PCR).The family history of asthma and history of allergic diseases were collected.2.THP-1 cells were induced into macrophages,miR-29b interference,miR-29b overexpression and normal control were induced by LV526,LV527 and NC virus infection.After 24 hours of culture,the cells were collected to detect the expression of STAT3 and B7H3 genes and proteins.3.It was verified that STAT3 was the target gene of miR-29b:after inoculating THP-1 cells and culturing with PMA with final concentration of 50ng/ml for 6 hours,the macrophages without PMA were cultured for 24 hours,then the macrophages infected by LV528,LV529 and NC virus were induced to form miR-29b interference,miR29b overexpression and normal control group.Luciferase analysis was performed at 48 hours to verify that STAT3 was the target gene of miR-29b.STAT3-3'UTR luciferase reporter gene plasmids were constructed and divided into three groups:"miR-29b+STAT3-3'UTR","miR-29b+STAT3-mut-3'UTR"and"miR-29b+luciferase empty load".4.Macrophages with different treatments were co-cultured with initial T cells for 3 days.The relative expressions of T-bet,GATA3 and ROR-γt were detected by Q-PCR.Result:1.The incidence of allergic disease in the acute attack group(68%)was higher than that in the other two groups(34.8%,33.3%),and the family history of asthma in the normal group(0%)was much lower than that in the other two groups(52%,60.9%).The difference was statistically significant(P<0.05).2.The expression of B7H3 in PBMC in acute attack group was higher than that in non-acute attack group and normal group.The expression of miR-29b in PBMC in normal group was significantly higher than that in non-acute attack group and acute attack group(P<0.0001).The expression of miR-29b in non-acute attack group was significantly higher than that in acute attack group(P=0.007).3.After silencing the expression of miR-29b,IL-4Rα,IL-4,IL-5,IL-13 and CD206 of macrophages increased significantly,while IFN-γdecreased,suggesting that miR-29b can promote the polarization of macrophages to M2.4.The overexpression of miR-29b,STAT3 and B7H3 gene and protein level in macrophages decreased,while the increase of miR-29b,STAT3 and B7H3 gene and protein expression was inhibited.5.There was a significant positive correlation between the expression of STAT3 and B7H3mRNA in macrophages(r=0.9737,P<0.0001).6.STAT3 is the target gene of miR-29b.7.Co-culture of macrophages with CD4^(+)T cells can promote the differentiation of primary T cells,namely Th 0 cells,into Th2,and the promoting effect of macrophages with downregulation of miR-29b is more obvious.Conclusion:The expression of miR-29b in PBMC of children with asthma is lower than that of normal children,while the expression of B7H3 is higher than that of normal children.It is speculated that miR-29b has a protective effect on children with asthma,while B7H3 aggravates the inflammatory response.Down-regulation of miR-29b,in macrophages can promote macrophages to M2 polarization,increase the expression of B7H3 and STAT3 in macrophages,make Th0 cells differentiate into Th2 cells,and aggravate the inflammatory response in patients with asthma.

四川盆地雷四^(3)亚段油气勘探方向

近年来,中石油和中石化在四川盆地雷口坡组雷四^(3)亚段的勘探陆续取得重要进展,展示了良好的勘探前景。笔者结合新钻探井和最新研究成果,从地层、沉积相、烃源岩、储层和成藏等多方面采用烃源对比、埋藏史及烃源岩热演化史、包裹体测温等方法对四川盆地雷四^(3)亚段的含油气地质条件进行了进一步分析,总结了其油气成藏特征,提出了有利勘探区带,以期有助于拓展四川盆地雷口坡组的勘探领域。研究结果表明:雷四^(3)亚段天然气以雷口坡组和须家河组烃源岩混合来源气为主;储集空间以粒间溶孔、晶间溶孔为主,属低孔低渗储层;雷四^(3)亚段气藏为2期成藏,烃源岩于中、晚侏罗世进入生烃高峰;雷四^(3)亚段具有构造和构造-岩性地层2种气藏类型,龙门山山前断褶带为构造气藏的有利勘探区,新津-邛崃斜坡带与梓潼-盐亭斜坡带是构造-岩性地层气藏的有利勘探区。

酮咯酸氨丁三醇口崩片处方优化及体外溶出度研究

目的优化酮咯酸氨丁三醇口崩片处方,并建立体外溶出度测定方法。方法以主观指标(口感、外观)、客观指标(硬度、崩解时间)为考察因素,采用模糊综合评分法联合L_(9)(3^(4))正交试验,对填充剂微晶纤维素(MCC)、甘露醇、崩解剂交联羧甲基纤维素钠(CCMC-Na)和硬脂酸镁用量进行优化,并验证处方工艺;采用高效液相色谱(HPLC)法测定酮咯酸氨丁三醇口崩片溶出度。结果酮咯酸氨丁三醇口崩片最佳处方为MCC 150 mg、甘露醇90 mg、CCMC-Na 10 mg、硬脂酸镁1.5 mg。制备的酮咯酸氨丁三醇口崩片口感良好,片面光洁,可压性强且硬度适中,崩解完全,且批间重复性良好。酮咯酸氨丁三醇质量浓度在1~100μg/mL范围内与峰面积线性良好(r=0.9999,n=5);平均回收率为98.76%,RSD为0.75%(n=9);精密度、稳定性、重复性试验结果的RSD均低于1.0%(n=6)。样品在10 min后即可溶解完全,溶出度均超过90%。结论优化的处方工艺简单、重复性好,可为酮咯酸氨丁三醇口崩片的工业化生产提供参考。

2-Gy whole-body irradiation significantly alters the balance of CD4^(+)CD25^(-) T effector cells and CD4^(+)CD25^(+)Foxp3^(+) T regulatory cells in mice

CD4^(+)CD25^(+) T regulatory(Treg)cells are critical in inducing and maintaining immunological self-tolerance as well as transplant tolerance.The effect of low doses of whole-body irradiation(WBI)on CD41CD251Foxp31 Treg cells has not been determined.The proportion,phenotypes and function of CD4^(+)CD25^(+) Treg cells were investigated 0.5,5 and 15 days after euthymic,thymectomized or allogeneic bone marrow transplanted C57BL/6 mice received 2-Gy c-rays of WBI.The 2-Gy WBI significantly enhanced the ratios of CD41CD251 Treg cells and CD4^(+)CD25^(+)Foxp3^(+) Treg cells to CD41 T cells in peripheral blood,lymph nodes,spleens and thymi of mice.The CD41CD251 Treg cells of the WBI-treated mice showed immunosuppressive activities on the immune response of CD4^(+)CD25^(+) T effector cells to alloantigens or mitogens as efficiently as the control mice.Furthermore,2-Gy c-ray WBI significantly increased the percentage of CD4^(+)CD25^(+)Foxp3^(+) Treg cells in the periphery of either thymectomized mice or allogeneic bone marrow transplanted mice.The in vitro assay showed that ionizing irradiation induced less cell death in CD4^(+)CD25^(+)Foxp3^(+) Treg cells than in CD4^(+)CD25^(+) T cells.Thus,a low dose of WBI could significantly enhance the level of functional CD41CD251Foxp31 Treg cells in the periphery of naive or immunized mice.The enhanced proportion of CD41CD251Foxp31 Treg cells in the periphery by a low dose of WBI may make hosts more susceptible to immune tolerance induction.

Alterations of peripheral CD4^(+)CD25^(+)Foxp3^(+)T regulatory cells in mice with STZ-induced diabetes

Complications arising from abnormal immune responses are the major causes of mortality and morbidity in diabetic patients.CD4^(+)CD25^(+)T regulatory cells(Tregs)play pivotal roles in controlling immune homeostasis,immunity and tolerance.The effect of hyperglycemia on CD4^(+)CD25^(+)Tregs has not yet been addressed.Here we used streptozotocin(STZ)-induced diabetic mice to study the effects of long-term hyperglycemia on CD4^(+)CD25^(+)Tregs in vivo.Four months after the onset of diabetes,the frequency of CD4^(+)CD25^(+)Foxp3^(+)T regulatory cells was significantly elevated in the spleen,peripheral blood lymphocytes(PBLs),peripheral lymph nodes(pLNs)and mesenteric LNs(mLNs).CD4^(+)CD25^(+)Tregs obtained from mice with diabetes displayed defective immunosuppressive functions and an activated/memory phenotype.Insulin administration rescued these changes in the CD4^(+)CD25^(+)Tregs of diabetic mice.The percentage of thymic CD4^(+)CD25^(+)naturally occurring Tregs(nTregs)and peripheral CD41Helios1Foxp31 nTregs were markedly enhanced in diabetic mice,indicating that thymic output contributed to the increased frequency of peripheral CD4^(+)CD25^(+)Tregs in diabetic mice.In an in vitro assay in which Tregs were induced fromCD4^(+)CD25^(+)T cells by transforming growth factor(TGF)-b,high glucose enhanced the efficiency of CD4^(+)CD25^(+)Foxp3^(+)T inducible Tregs(iTregs)induction.In addition,CD4^(+)CD25^(+)T cells from diabetic mice were more susceptible to CD4^(+)CD25^(+)Foxp3^(+)TiTreg differentiation than those cells from control mice.These data,together with the enhanced frequency of CD4^(+)CD25^(+)Foxp3^(+)T iTregs in the periphery of mice with diabetes,indicate that enhanced CD4^(+)CD25^(+)Foxp3^(+)T iTreg induction also contributes to a peripheral increase in CD4^(+)CD25^(+)Tregs in diabetic mice.Our data show that hyperglycemia may alter the frequency of CD4^(+)CD25^(+)Foxp3^(+)T Tregs in mice,which may result in late-state immune dysfunction in patients with diabetes.

三味板蓝根颗粒浸膏制备工艺改良研究

目的改良医院制剂三味板蓝根颗粒浸膏中间体的制备工艺。方法采用L_(9)(3^(4))正交试验优选浸膏提取工艺的关键参数,再以单因素和Box-Behnken设计-响应面法优化浸膏醇沉工艺。结果正交试验优选的浸膏最佳提取工艺为:以10倍量水(初次浸泡0.5 h),沸腾提取3次,每次提取1.5 h。经单因素和响应面法优化浸膏最佳醇沉工艺为:将药液浓缩至相对密度为1.15(80℃),以95%乙醇将药液的终点醇浓度调至65%,醇沉静置时间48 h。结论采用L_(9)(3^(4))正交试验联合Box-Behnken设计-响应面法优化医院制剂三味板蓝根颗粒的提取和醇沉工艺,达到预期效果,可为实际扩大化生产提供参考。

基于层次分析法的正交试验优选坐浴安散提取工艺

目的优选坐浴安散的提取工艺。方法以醇提物中大黄酸、大黄素、大黄酚含量和醇提干膏得率为综合评价指标,采用层次分析(AHP)法确定各指标权重,结合L_(9)(3^(4))正交试验考察乙醇体积分数、溶剂用量、提取时间、提取次数对醇提工艺的影响;以水提物中盐酸小檗碱、苦参碱、氧化苦参碱含量和水提干膏得率为综合评价指标,采用AHP法确定各指标权重,结合L_(9)(3^(4))正交试验考察加水量、提取时间、提取次数对水提工艺的影响。结果优选的醇提工艺为加8倍量70%乙醇,提取3次,每次40 min;水提工艺为加12倍量水,提取3次,每次1.0 h。结论AHP法结合L9(34)正交试验优选的提取工艺合理、可行,能较好地保证坐浴安散的质量。

小儿清热颗粒指标成分测定和水提工艺研究

目的建立小儿清热颗粒指标成分测定方法,并优选最佳水提工艺。方法采用高效液相色谱法测定羟基红花黄色素A含量,以加水量、提取次数、提取时间为考察因素,以提取液干膏得率、乙酸乙酯萃取部位得率和羟基红花黄色素A转移率为评价指标,设计L_(9)(3^(4))正交试验以优选最佳水提工艺。结果羟基红花黄色素A对照品溶液质量浓度在23.44~195.34 μg/mL范围内与峰面积线性关系良好(r=0.9996,n=6),精密度、稳定性和重复性试验结果的RSD均低于2.0%,平均加样回收率为100.69%,RSD为0.81%(n=6)。最佳水提工艺为加10倍水,提取3次,每次0.5 h。结论所建立的指标成分含量测定方法及优选的水提工艺稳定可靠、重复性好,可为藏药小儿清热颗粒的进一步开发奠定基础。

Conversion of effector CD4^(+)T cells to a CD8^(+)MHC Ⅱ-recognizing lineage

CD4^(+)and CD8^(+)T cells are dichotomous lineages in adaptive immunity.While conventionally viewed as distinct fates that are fixed after thymic development,accumulating evidence indicates that these two populations can exhibit significant lineage plasticity,particularly upon TCR-mediated activation.We define a novel CD4^(-)CD8αβ^(+)MHC Ⅱ-recognizing population generated by lineage conversion from effector CD4^(+)T cells.CD4-CD8αβ^(+)effector T cells downregulated the expression of T helper cell-associated costimulatory molecules and inaeased the expression of cytotoxic T lymphocyte-associated cytotoxic molecules.This shift in functional potential corresponded with a CD8^(+)-lineage skewed transcriptional profile.TCRβ repertoire sequencing and in vivo genetic lineage tracing in acutely infected wild-type mice demonstrated that CD4^(-)CD8αβ^(+)effector T cells arise from fundamental lineage reprogramming of bona fide effector CD4^(+)T cells.Impairing autophagy via functional deletion of the initiating kinase Vps34 or the downstream enzyme Atg7 enhanced the generation of this cell population.These findings suggest that effector CD4^(+)T cells can exhibit a previously unreported degree of skewing towards the CD8^(+)T cell lineage,which may point towards a novel direction for HIV vaccine design.

正交试验法优选抗早颗粒提取及成型工艺

目的优选抗早颗粒的提取及成型工艺。方法以盐酸小檗碱含量和干膏得率的综合评分为评价指标,考察提取时间、提取次数、加水量,采用L_(9)(3^(4))正交试验法优选水提工艺;以颗粒得率和水分为评价指标,考察雾化压力、喷雾速率、物料温度,采用L_(9)(3^(4))正交试验优选流化床制粒工艺。结果提取次数对水提工艺影响显著,最佳水提工艺为加8倍量水,提取2次,每次40 min。雾化压力和喷雾速率对流化床制粒工艺的颗粒得率影响显著,最佳成型工艺为喷雾速率20 r/min,雾化压力0.14 MPa,物料温度55℃。结论该优选工艺稳定性好,可用于抗早颗粒的工业化生产。

多抗乙素对小鼠S_(180)肉瘤中免疫活性细胞浸润的影响

曾经证明多抗乙素有明显的抑瘤作用。本文以小鼠实验性S_(180)肉瘤为模型观察了多抗乙素对肿瘤组织内和肿瘤周围Mφ、L_3T_4^- 及Lyt_2^-细胞的影响,同时观察了肿瘤坏死组织周围中性粒细胞浸润及小血管内血栓形成的改变。结果提示:Pb治疗组,肿瘤周围及肿瘤内浸润的L_3T_4^-和Lyt_2^-细胞显著增多,说明Pb可诱导肿瘤组织中淋巴细胞浸润;肿瘤组织的坏死灶周围明显充血、小血管血栓形成显著增加,并有带状中性粒细胞浸润。这些改变与TNF介导的肿瘤组织损伤相符,提示Pb的抑瘤作用主要是通过免疫活性细胞产生的TNF实现的。

基于信息熵赋权法的正交试验优选细辛-干姜药对水提取工艺

目的优选细辛-干姜药对的水提取工艺。方法以浸泡时间、加水量、提取时间、提取次数为考察因素,以6-姜酚、芝麻脂素、细辛脂素、6-姜烯酚的含量和干浸膏得率为评价指标,以信息熵赋权法确定评价指标的权重系数,采用L_(9)(3^(4))正交试验优选细辛-干姜药对的最佳水提取工艺。结果确定6-姜酚、芝麻脂素、细辛脂素、6-姜烯酚的含量和干浸膏得率的权重系数分别为0.1537,0.1562,0.1662,0.2544,0.2695;优选的最佳水提取工艺为加6倍量水,浸泡1.0 h,提取3次,每次2.0 h。按此工艺制备的样品中,6-姜酚、芝麻脂素、细辛脂素、6-姜烯酚的含量分别为0.1901,0.0916,0.1774,0.1116μg/mL,干浸膏得率为30.91%(n=3)。结论优选的提取工艺稳定、可行,可为细辛-干姜药对的后续研究提供参考。

正交试验优选通癃颗粒提取工艺

目的优选通癃颗粒的提取工艺。方法以加水量、提取时间、提取次数为考察因素,以毛蕊花糖苷、马钱苷和莫诺苷含量及干膏得率为评价指标,采用L_(9)(3^(4))正交试验优选通癃颗粒的提取工艺。结果最佳提取工艺为加8倍量水,提取3次,每次1.0 h。在此工艺下制备的样品中,毛蕊花糖苷、马钱苷和莫诺苷的平均含量分别为24.78μg/mL和53.92μg/mL,平均干膏得率为17.21%(n=3)。结论该提取工艺稳定、可行,可为通癃颗粒的工业化生产提供参考。

CD56^brightCD25^＋ NK cells are preferentially recruited to the maternal/fetal interface in early human pregnancy

Decidual natural killer （dNK） cells are believed to be critical for maintaining maternal/fetal tolerance and regulating placental vascular remodeling based upon their abundance and unique phenotype during early pregnancy. However, the mechanism for how the dNK cells play such important roles in successful pregnancy remains undefined. Here, we identified a subtype of dNK cells characterized as having a CD3-CD56^brightCD25^＋ phenotype. We found that CD56^brightCD25^＋ NK cells preferentially localize to the maternal/fetal interface during early human pregnancy. CD25^＋ dNK cells account for approximately 75% of CD25-expressing decidual immune cells （DICs）. However, less than 5% of CD25-positive peripheral blood mononuclear cells are CD25^＋ NK cells. Furthermore, CD25^＋ and CD25^- dNK cells exhibit distinct phenotypes： CD25^＋ dNK cells display a more activated phenotype and greater cytokine-secreting capacity. Interestingly, coculture of peripheral NK （pNK） cells with primary trophoblasts upregulates the percentage of CD25-expressing pNK cells, resulting in increased expression of activation markers and cytokine production by pNK cells. In addition, we demonstrated that the CXCL12/CXCR4 axis is crucial for the recruitment of CD25^＋ dNK cells and contributes to the accumulation of CD3^-CD56^brightCD25^＋ dNK cells at the maternal/fetal interface. Thus, our data reveal that the crosstalk between trophoblasts and pNK cells leads to the accumulation of CD3^-CD56^brightCD25^＋ dNK cells, which exert a regulating effect at the maternal/fetal interface.