超声引导高温蒸馏水加干扰素瘤内注射治疗肝癌的研究

目的探讨超声引导瘤内注射高温蒸馏水及干扰素治疗肝癌的安全性、疗效及免疫学变化。 方法对36例肝癌患者57个肿瘤，在超声引导下经皮肿瘤穿刺注射高温蒸馏水（90℃）加干扰素治疗，观察治疗效果及免疫学变化，并与CT、彩色多普勒超声及实验室检查情况相对照。 结果超声见肿瘤缩小32个（56．1％）；彩色多普勒超声肿瘤内血流消失46个（80．7％）；肿瘤内血流明显减少8个（14％）；57个肿瘤均行超声引导下活检，病理结果为大片状坏死组织，肿瘤完全坏死49个（86．0％），肿瘤不完全坏死8个（14％）。36例治疗后1个月行CT检查，肿瘤体积缩小31个（54．4％），增强CT显示肿瘤区内无明显强化，呈低密度；血清免疫球蛋白IgA、IgG、IgM和补体C3、C4及T淋巴细胞转化试验均有明显升高。 结论应用超声引导下高温蒸馏水加干扰素瘤内注射治疗肝癌效果显著，安全性高，能提高免疫功能，是一种值得深入探讨的治疗方法。     

超声造影引导下经皮经肝注射无水乙醇治疗肝癌的临床观察

目的：探讨超声造影引导下经皮经肝注射无水乙醇治疗肝癌的可行性和有效性 方法：对我院2013年3月至2015年5月,病理证实的原发性肝癌患者86例,共计107个病灶,行超声造影引导下经皮经肝无水乙醇注射治疗肝癌,并行超声造影对其疗效进行观察。 结果：107个病灶经超声造影引导下行经皮经肝无水乙醇注射治疗后,经超声造影评价,局部消融成功率达到100%,随访时间6月至2年,肿瘤局部进展率5.6%（6/107）,未发现严重消融相关并发症。 结论：超声造影引导下行经皮经肝无水乙醇注射治疗肝癌,可以有效的杀灭肿瘤组织,是一种临床可行有效的介入治疗肿瘤的方法。     

超声引导瘤内注射无水酒精治疗子宫肌瘤的临床研究

子宫肌瘤治疗方法有手术、保守和介入治疗等[1],某些治疗可能存在严重并发症的风险,非血管途径有超声引导注射无水酒精、射频、微波、冷冻、高能聚焦超声热疗等方法[2]也存在依赖于仪器设备的性能;作者采用经阴道超声引导子宫肌瘤瘤内注射无水酒精(浓度≥99.7%)治疗31例,现报道如下.     

超声引导瘤内注射亲细胞非均质分子脂质治疗肝癌新方法的临床应用

目的探讨超声引导下不饱和脂肪酸类药物--亲细胞非均质分子脂质(CHML)肝癌瘤内注射临床应用效果.方法总结经自动活检病理证实肝细胞肝癌15例注射CHML治疗后的临床疗效,观察治疗前后AFP、CDFI和CT局部肿瘤的改变,并计算半年生存率.结果15例肝细胞肝癌(22结节)用CHML瘤内注射后,瘤体体积均无增大或者明显减小,8例(8/9)AFP下降,7例行CT复查瘤体缩小,增强扫描无强化(7/7),9例注射CHML 2～3次后穿刺病理复查发现肿瘤细胞变性、坏死伴瘤周纤维组织增生,血管内血栓形成,但无血管扩张及增生.CHML注射治疗前后肝、肾、肺功能无明显改变(P＞0.05).结论CHML作为一种局部硬化剂具有明显的抗肿瘤作用,近期临床效果较好.其作用机制可能为造成肿瘤血管内血栓形成,导致肿瘤血供障碍,初步观察认为,CHML与无水乙醇混合应用效果可更佳.     

超声引导肝癌介入性治疗进展

当前超声引导下肝癌的间质介入治疗方法有几十种,根据其方式不同可分为局部液体性制剂注射治疗和局部间质热疗或冷冻治疗。本文仅就几种临床较常用的方法综述如下：１、局部注射治疗　局部注射治疗是在超声引导下将注射剂注入瘤体内,通过化学或物理效应使瘤体坏死。目前使用的注射剂包括无水乙醇、醋酸溶液、热盐水、化疗药物、ＯＫ－４３２及放射性同位素等。经皮瘤内无水乙醇注射治疗（ＰｅｒｃｕｔａｎｅｏｕｓＥｔｈａｎｏｌＩｎｊｅｃｔｉｏｎＴｈｅｒａｐｙ,ＰＥＩＴ）,具有灭活效果好、毒副作用小、方法简便实用、价廉等优点。自…     

超声引导经皮穿刺无水乙醇注射治疗肝癌的临床应用价值

目的探讨超声引导经皮穿刺无水乙醇注射治疗肝癌的临床应用价值。方法对58例肝癌患者共70个结节在超声引导下细针穿刺经皮瘤内注射无水乙醇进行硬化治疗。肿瘤直径1～4cm、每个瘤体内注射2～6次、注射剂量6～60ml。结果58例在注射2～6次后，总有效率为56．40％。治疗后12个瘤灶消失；34个缩小；24个未增大。直径缩小或不变的肿瘤CT增强扫描动脉期无强化。原血清血甲胎蛋白（AFP）值升高者显著下降。经组织学检查的病灶证实完全坏死。注射过的瘤灶无局部复发。结论超声引导经皮穿刺无水乙醇注射治疗肝癌是一种简单易行、安全和较为有效的方法。     

超声引导下射频消融治疗肝癌的临床价值

目的探讨超声引导下射频消融(RFA)治疗肝癌的疗效。方法35例肝癌49个病灶行超声引导下RFA治疗,对治疗前后病灶的超声影像、超声造影、螺旋CT检查、肝功能、AFP变化以及机体免疫功能的影响等进行综合研究以评价疗效。结果RFA后5min,超声造影,肿块中83.7%(41/49)各个时相无增强,即回声缺失;1周后彩超示肿块中87.8%(43/49)血流信号明显减少或消失;1个月后螺旋CT扫描肿块中81.6%(40/49)大小无明显变化,但肿块内无强化。肝功能有不同程度改善,甲胎蛋白下降,CD4 、CD4 /CD8 明显升高,sIL-2R明显降低。结论超声引导下射频消融治疗肝癌是有效的微创治疗手段,其为操作简便、损伤小、安全性高的有效方法。     

超声引导下射频消融治疗肝癌的临床价值

目的：探讨超声引导下多极射频介入治疗肝癌的临床价值。方法：19例肝癌患者26枚结节在超声引导下进行射频消融治疗，采取由深到浅，多点多部位，重叠消融的治疗原因，结果：12个结节（46．2％）缩小，肿瘤回声均增强16个（61．5％）血流信号减少或消失；病检8例阴性（8／12）；10例甲胎蛋白下降（10／13）；7例（7／13）肝功能改善，结论：超声引导下多电极射频治疗肝癌是一种疗效确切，操作简便，损伤小，安全性高的有效方法。     

超声引导下瘤体内注入超声造影剂和P53基因治疗大鼠肝癌的初步研究

目的　探讨超声引导下将超声造影剂和人野生型 p5 3基因直接注入瘤体内对大鼠肝癌基因表达的影响。方法　采用免疫抑制法建立大鼠原位肝癌模型。 2 4只实验Wistar大鼠随机分成 4组。第 1组 :超声引导下将基因直接注入瘤体内并不用超声照射 ;第 2组 :注入基因后用超声照射瘤体 ;第 3组 :注入造影剂和p5 3基因后不行超声照射 ;第 4组 :用超声照射注入造影剂和基因的瘤体。 48h后用半定量逆转录 聚合酶链式反应 (RT PCR)测定肝癌细胞内 p5 3mRNA的表达情况。 结果　超声引导下可准确地将基因注入肝癌内 ,4组肝癌细胞内均有 p5 3mRNA的表达 ,第 4组的表达量最高 ,明显高于其他三组 ( P <0 .0 0 1)。第 2组的基因表达量次之 ,高于另外两组 (P <0 .0 5 )。第 1、3组间在外源基因表达上的差异没有显著性意义 (P >0 .0 5 )。结论　超声引导下瘤体内注射超声造影剂和p5 3基因后用超声照射 ,既可有效地控制肝癌基因治疗的靶向性 ,又能提高外源基因的表达量。     

肝星状细胞与肝癌的研究进展

原发性肝癌(以下简称肝癌)是我国常见的恶性肿瘤之一,尽管在最近几十年对其的研究取得了可喜的进展,但肝癌的5年生存率仍只有5%左右。肝癌是在环境序贯作用下由一群正常细胞转化而来的遗传不均一的细胞群,肝癌细胞和间质成分构成的微环境在肝癌研究中日益被重视。正确认识肝癌微环境并把握肝癌细胞与间质相互作用的机制,对于认识肝癌起始、演进和转移的全过程具有重要意义。在我国,大多数肝癌患者合并乙型肝炎     

超声介入无水酒精量化治疗肝癌的临床评价

目的：评价无水酒精量化注射治疗肝癌（HCC）的临床应用价值。方法：对122例经病理证实的肝癌在超声引导下量化注射无水酒精（PEI），其中肿瘤直径＞3cm58例、肿瘤直径≤3cm64例。注射量按回归方程Y＝2．885X（当肿瘤直径≤5cm时），Y＝1．805X（当肿瘤直径＞5cm时）计算，式中X为肿瘤最大直径（cm），Y为注射酒精量（ml），每3－5天注射一次，4－10次为一疗程，＞5cm的肿瘤结节可以10－20次为一疗程。随访12－48个月，观察1、2、3、4年的生存率、原位与异位复发率。结果：122例HCC经量化无水酒精注射治疗后，B超示62．5％肿瘤直径缩小，平均直径从3．8cm降为2．9cm；25．0％肿瘤结节周边见高回声环，其外包绕低回声暗晕；12．5％肿瘤结节消失。肿瘤直径≤3cm组1、2、3、4年生存率分别为94％、85％、72％与63％，高于直径＞3cm组的84％、64％、58％与52％，而原位复发率前者分别为0％、2．1％、5．1％与4．2％，明显低于后者的6．9％、7．1％、24．2％与28．6％；异位复发率肿瘤直径≤3cm组分别为9．0％、10．4％、12．8％与20．8％，也明显低于直径＞3cm组的15．5％、21．4％、36．4％与52．4％。122例先后进行1221次PEI，除1例出现黑便，4例出现黄疸，经一般治疗1－2周后恢复外，其他无大出血和严重心、肝、肾功能损害等并发症发生。结论：超声介入量化注射无水酒精治疗HCC疗效可靠，尤其是肿瘤直径≤3cm的疗效更佳。它还具有副作用小、操作方便、相对低廉等优点，有较高的临床应用价值，值得深入研究。     

Rapid Effector Function in CD8+ Memory T Cells

The nature of the CD8⁺ T cells that underlie antiviral protective immunological memory in vivo is unclear. We have characterized peptide-specific CD8⁺ T lymphocytes directly ex vivo from peripheral blood in humans with past exposure to influenza virus, using single cell interferon γ (IFN-γ) release as a measure of effector function. In individuals in the memory state with respect to influenza virus infection, unrestimulated antigen-specific CD8⁺ T cells displayed IFN-γ release within 6 h of antigen contact, identifying a population of memory CD8⁺ T cells that exhibit effector function without needing to divide and differentiate over several days. We have quantified circulating CD8⁺ effector T cells specific for six different MHC class I–restricted influenza virus epitopes. Enumeration of these CD8⁺ T cells gives frequencies of peptide-specific T cells that correlate with, but are in general severalfold higher than, CTL precursor frequencies derived from limiting dilution analysis, indicating that this novel population of memory CD8⁺ T cells has hitherto been undetected by standard means. The phenotype of these cells, which persist at a low frequency long after recovery from an acute viral infection, suggests that they play a role in protective immunological memory.After recovery from an acute viral infection, the frequency of antigen-specific CD8⁺ T cells is too low to permit direct analysis. Instead, assays have used antigen- experienced T cells that have been expanded by in vitro restimulation with cognate antigen, a process that may introduce quantitative and qualitative biases, particularly with respect to the activation state of the cell. It has therefore been difficult to establish, for viral infections in humans, the phenotype of antigen-specific memory T cells in their natural state. In particular, it remains uncertain whether antiviral protective immunological memory is subserved by long-lived quiescent T cells (1–3) that must divide and differentiate over several days to become effectors, or by circulating effector T cells continuously activated either by persisting antigen or by cross-reactive environmental antigens (4, 5).We have studied individuals previously exposed to influenza virus but without acute clinical influenza, that is, individuals in the memory state with respect to influenza virus infection. Influenza infection is well suited for the study of CD8⁺ T cell memory. First, CD8⁺ CTLs are crucial for host defense against influenza virus; they are important in the clearance of intranasal virus (6, 7) and, since CTLs recognize the relatively conserved internal viral proteins (8, 9), they cross-react between viruses of different strains which evade neutralizing antibody through variation in surface glycoproteins (10, 11). Second, influenza virus–specific CTLs secrete IFN-γ which has direct effects on virus replication in infected cells (12) and may be more important in vivo for protection against influenza virus than perforin- or fas-mediated lysis (13). Examination of IFN-γ secretion by antigen-specific CD8⁺ T cells is therefore expected to be of at least as much relevance to protection as conventional measurements of lytic activity.We have applied a sensitive enzyme-linked immunospot (ELISPOT)¹ assay for single cell IFN-γ secretion in a novel way to detect low frequencies of uncultured influenza peptide–specific CD8⁺ T lymphocytes freshly isolated from peripheral blood. The ELISPOT assay detects secreted cytokine molecules in the immediate vicinity of the cell from which they are derived, while still at a relatively high concentration; each spot in the read-out represents a ‘footprint'' of the original cytokine-producing cell. Quantitation of these IFN-γ spot-forming cells (SFCs) by this technique is highly sensitive; for influenza virus–specific CD8⁺ CTL lines and bulk cultures, the ELISPOT assay is an order of magnitude more sensitive than the ⁵¹Cr-release cytotoxicity assay for detecting low numbers of peptide-specific CTLs (data not shown). We have exploited this enhanced sensitivity to demonstrate the presence of circulating influenza virus–specific CD8⁺ memory T cells capable of rapid effector function, long after exposure to the virus.     

Viral Immune Evasion Due to Persistence of Activated T Cells Without Effector Function

We examined the regulation of virus-specific CD8 T cell responses during chronic lymphocytic choriomeningitis virus (LCMV) infection of mice. Our study shows that within the same persistently infected host, different mechanisms can operate to silence antiviral T cell responses; CD8 T cells specific to one dominant viral epitope were deleted, whereas CD8 T cells responding to another dominant epitope persisted indefinitely. These virus-specific CD8 T cells expressed activation markers (CD69^hi, CD44^hi, CD62L^lo) and proliferated in vivo but were unable to elaborate any antiviral effector functions. This unresponsive phenotype was more pronounced under conditions of CD4 T cell deficiency, highlighting the importance of CD8– CD4 T cell collaboration in controlling persistent infections. Importantly, in the presence of CD4 T cell help, adequate CD8 effector activity was maintained and the chronic viral infection eventually resolved. The persistence of activated virus-specific CD8 T cells without effector function reveals a novel mechanism for silencing antiviral immune responses and also offers new possibilities for enhancing CD8 T cell immunity in chronically infected hosts.     

B-Cell-Delivered Gene Therapy Induces Functional T Regulatory Cells and Leads to a Loss of Antigen-Specific Effector Cells

Previous reports have shown that B-cell-mediated gene therapy can induce tolerance in several animal models for autoimmune diseases and inhibitory antibody formation in hemophilia A mice. We know from our previous work that the induction of tolerance following B-cell therapy is dependent upon CD25⁺ regulatory T cells (Tregs). To extend these studies and identify the effects of this gene therapy protocol on the target CD4 T cells, we have adapted in vitro suppression assays using Tregs isolated from treated and control mice. Using carboxyfluorescein succinimidyl ester (CFSE) dilution as a measure of T-cell responsiveness to FVIII, we show that CD25⁺ Tregs from treated mice are more suppressive than those from control animals. To monitor the induction of antigen-specific Tregs, we repeated these studies in ovalbumin (OVA) peptide-specific DO11.10 T-cell receptor (TCR) transgenic mice. Tregs from DO11.10 mice treated with a tolerogenic OVA–Ig construct are better than polyclonal Tregs at suppressing the proliferation of responder cells stimulated with OVA peptide 323–339 (pOVA). Furthermore, we show that following B-cell therapy, there is an increase in antigen-specific FoxP3⁺ Tregs, and there is also a distinct decrease in antigen-specific CD4⁺ effector T cells. These changes in the lymphocyte population shift the balance away from effector function toward a tolerogenic phenotype.     

超声引导下经皮门静脉穿刺注药治疗肝癌的应用

目的　研究超声引导下经皮门静脉穿刺注药治疗肝癌的应用价值 ,旨在提高肝动脉栓塞化疗的疗效和降低肝癌合并门静脉转移率。方法　对 14例不能手术的肝癌患者在行肝动脉栓塞治疗 1周后 ,超声引导下经皮门静脉穿刺注药化疗 ,化疗药为顺铂 30 mg,5 - Fu 5 g。结果　 (1)治疗后所有患者的肿块均有明显缩小 ,其中3例缩小 >4 0 % ;(2 )肝癌合并门静脉癌栓患者 6例 ,门静脉癌栓消失率为 16 .7% (1/ 6 ) ,缩小率为 5 0 % (3/ 6 ) ,无变化 33% (2 / 6 ) ;(3)治疗后 10例原发性肝癌 AFP升高者 ,7例下降超过 5 0 % ,2例无变化 ,1例升高 ;(4)患者腹胀减轻 ,腹水减少。结论　超声引导下经皮门静脉穿刺注药治疗 ,确实有效 ,方法简便、安全 ,无并发症 ,作为肝癌综合治疗的方法之一 ,值得进一步探索。     

热处理小鼠H22肝癌细胞的抗肿瘤作用

目的：比较肿瘤细胞体外经不同温度时间热处理后的抗肿瘤效应。方法：采用普通水浴加热方法将小鼠Ｈ２２肝癌细胞经一定温度时间处理后制成瘤苗，免疫ＩＣＲ小鼠，然后腹腔移植Ｈ２２细胞，观察小鼠的平均存活天数，免疫小鼠脾细胞体外对Ｈ２２肝癌细胞的杀伤作用。结果：６５℃／３０分钟水浴加热处理能完全杀灭Ｈ２２细胞；６５℃／３０分钟热处理制备的瘤苗能显著延长荷瘤小鼠的存活时间（Ｐ＜０．０１）；并且该瘤苗免疫小鼠的脾细胞杀伤Ｈ２２细胞作用显著高于荷瘤小鼠及正常小鼠（Ｐ＜０．０１）。结论：采用水浴加热方法制备的肿瘤瘤苗能增强机体的抗肿瘤作用，６５℃／３０分钟为水浴方法制备瘤苗的适宜条件。     

肝癌细胞HepG2和正常肝细胞LO2放射敏感性体外实验研究

目的：探讨体外培养的人肝癌细胞株HepG2和人正常肝细胞株LO2的放射敏感性。方法采用6-MVX线分次照射细胞,进行克隆形成实验,流式细胞仪测细胞凋亡。结果 HepG2的细胞存活曲线较陡峭,细胞凋亡率升高幅度较达。结论 HepG2与LO2有不同的放射生物学特性,相比而言HepG2的放射敏感性较高。     

Concurrent Generation of Effector and Central Memory CD8 T Cells during Vaccinia Virus Infection

It is generally thought that during the contraction phase of an acute anti-viral T cell reponse, the effector T cells that escape activation-induced cell death eventually differentiate into central memory T cells over the next several weeks. Here we report that antigen-specific CD8T cells with the phenotype and function of central memory cells develop concomitantly with effector T cells during vaccinia virus (vv) infection. As soon as 5 days after an intraperitoneal infection with vv, we could identify a subset of CD44^hi and CD62L⁺ vv-specific CD8 T cells in the peritoneal exudate lymphocytes. This population constituted approximately 10% of all antigen-specific T cells and like central memory T cells, they also expressed high levels of CCR7 and IL-7R but expressed little granzyme B. Importantly, upon adoptive transfer into naïve congenic hosts, CD62L⁺, but not CD62L⁻ CD8 T cells were able to expand and mediate a rapid recall response to a new vv challenge initiated 6 weeks after transfer, confirming that the CD62L⁺ vv-specific CD8 T cells are bonafide memory cells. Our results are thus consistent with the branched differentiation model, where effector and memory cells develop simultaneously. These results are likely to have implications in the context of vaccine design, particularly those based on vaccinia virus recombinants.