食管鳞癌组织中B7-H4、CD3表达变化及意义

目的探讨B7-H4、CD3在食管鳞癌发生、发展中的作用。方法采用免疫组化SP法检测30例食管鳞癌组织（食管癌组）和20例癌旁正常食管黏膜组织（对照组）中B7-H4、CD3蛋白表达,采用等级相关方法分析B7-H4和CD3表达的相关性。结果食管癌组B7-H4阳性表达率高于对照组,CD3阳性表达率低于对照组（P均〈0.05）。不同临床分期及淋巴结转移情况者食管癌组织中B7-H4表达差异有统计学意义（P均〈0.05）。食管癌组织中B7-H4与CD3蛋白的表达呈负相关关系（r=-0.467,P〈0.05）。结论食管鳞癌组织中B7-H4呈高表达,CD3呈低表达,二者与食管鳞癌的发生发展及侵袭转移有关。     

食管癌组织中p53、HPV的表达变化及意义

目的观察食管癌组织中p53、人乳头瘤病毒（HPV）的表达变化,并探讨其意义。方法取45例食管癌患者肿瘤组织组织（食管癌组）、20例食道良性肿瘤患者肿瘤组织（良性肿瘤组）、20例食管正常者的食管组织（对照组）,采用ELISA法检测其中p53蛋白,采用新柏式液基细胞学技术（TCT）检测HPV。结果食管癌组44例p53表达阳性、40例HPV阳性,良性肿瘤组分别为2、2例,对照组分别为1、1例,食管癌组p53、HPV阳性表达率与良性肿瘤组和正常组相比P均〈0.01,良性肿瘤组与对照组相比P〉0.05。食管癌组织中HPV、p53表达呈正相关（r=0.429,P〈0.05）。结论食管癌组织中p53、HPV表达升高。检测HPV、p53有助于食管癌早期诊断。     

HPV16 E6/E7基因及蛋白表达与宫颈癌的相关性研究

目的研究人乳头状瘤病毒16型（HPV16）E6／E7基因及其蛋白表达在宫颈疾病及其癌变中的作用。方法运用PCR技术检测51例宫颈癌（癌症组）、20例富颈上皮瘤变（CIN）Ⅱ～Ⅲ级（CIN组）、20例宫颈炎（炎症组）患者病变组织中HPV16 E6／E7基因，并运用免疫组化SP法检测癌症组癌组织中HPV16E6、E7的表达情况。结果癌症组、CIN组、炎症组HPV16 E6检出率分别为5％、35％、45％．后两者明显高于前者（P〈0．05），HPV16 E7检出率分别为65％、75％、68．6％，P均〉0．05；癌症组45例HPV16 E6和42例E7蛋白阳性表达（88．2％、82．3％）。HPV16 E6蛋白表达与临床分期、肿瘤分化程度和淋巴结有无转移均无相关性（P〉0．05），HPV16 E7蛋白表达与临床分期和淋巴结有无转移相关（P〈0．05），与肿瘤分化程度无相关性（P〉0．05）。结论HPV16 E6／E7基因与宫颈疾病及其癌变的关系密切。     

非小细胞肺癌组织中AQP1、nm23-H1的表达变化及意义

目的观察非小细胞肺癌组织中AQP1、nm23-H1蛋白的表达变化,并探讨其意义。方法采用免疫组化SP法检测123例非小细胞肺癌患者（观察组）和12例非肺癌受检者（对照组,包括7例肺炎性假瘤和5例正常受检者）肺组织中的AQP1、nm23-H1蛋白。结果观察组65例nm23-H1表达阳性,对照组1例;观察组77例AQP1阳性率,对照组2例。两组AQP1、nm23-H1阳性表达率相比,P均〈0.05。观察组AQP1表达与临床分期、病理分化程度、淋巴结转移、患者生存期有关（P均〈0.05）;nm23-H1蛋白表达仅与淋巴结转移相关（P〈0.05）。观察组AQP1和nm23-H1的表达呈正相关（P〈0.05）。结论观察非小细胞肺癌组织中AQP1、nm23-H1蛋白表达上调。二者联合检测可作为判断非小细胞肺癌转移潜能的指标     

年轻宫颈癌患者癌组织HPV16检测结果分析

采用PCR技术对31例年龄≤35岁的宫颈癌患者（年轻组）和54例年龄〉35岁的宫颈癌患者（对照组）的癌组织进行人乳头瘤病毒（HPV）16检测。结果显示，年轻组HPV16阳性者17例（54.8％），对照组阳性者13例（24.1％），两组相比，P〈0．05。提示宫颈癌发病年轻化与HPV16感染有关。     

贵州地区宫颈上皮内瘤变和宫颈浸润癌患者HPV感染状态及其亚型分布

目的观察贵州地区宫颈上皮内瘤变（CIN）和宫颈浸润癌患者人乳头状瘤病毒（HPV）的感染状态及其亚型分布。方法采用导流杂交技术对30例CIN（CIN组）、33例宫颈浸润癌患者（宫颈癌组）、60例自愿接受宫颈HPV感染筛查的妇女（对照组）进行HPV分型检测,所有受检者均来自贵州地区。结果 CIN组HPV阳性13例（43.33%）,宫颈癌组19例（57.58%）,对照组5例（8.33%）。CIN组、宫颈癌组与对照组比较,P均〈0.01。CIN组共检出7种亚型HPV,均为高危型（HR-HPV）,无多型HPV感染（M-HPV）。其中HPV16 7例,HPV31、52各2例,HPV18、33、53、58各1例。宫颈癌组检出5种亚型,亦均为HR-HPV,M-HPV3例。其中HPV16 11例,HPV534例,HPV58 3例,HPV18、59各1例。对照组检出7种亚型,HR-HPV中的HPV16、18、31、39、53、58和低危型HPV的HPV6各1例;M-HPV 1例。CIN组、宫颈癌组HPV16感染率明显高于对照组,P均〈0.05。HR-HPV感染与CIN（OR=8.412,95%CI为2.62~26.99）和宫颈浸润癌（OR=14.929,95%CI为4.74~46.98）紧密相关,且主要与HPV16感染有关（CIN：OR=17.957,95%CI为2.09~154.15;宫颈浸润癌：OR=29.500,95%CI为3.60~242.07）,P均〈0.05。结论贵州地区CIN和宫颈浸润癌与HR-HPV感染密切相关,HPV16为主要感染亚型。     

宫颈癌组织中SCCA、p53蛋白的表达变化及意义

目的观察宫颈癌组织中鳞状细胞癌抗原（SCCA）、p53蛋白的表达变化,并探讨其意义。方法宫颈癌患者45例（肿瘤组）,行宫颈癌根治术;子宫肌瘤患者20例（良性瘤组）,均行子宫肌瘤切除术,两组术中留取肿瘤组织,另取良性瘤组瘤旁2 cm以上正常组织作对照（对照组）。采用电化学发光法检测宫颈癌组织、子宫肌瘤组织及正常子宫组织中的SCCA;采用ELISA法对p53蛋白进行检测。结果肿瘤组SACC阳性23例,p53蛋白阳性21例,良性瘤组分别为3、2例,对照组分别为1、0例。肿瘤组与良性瘤组、对照组相比,P均〈0.01。肿瘤组SACC阳性的23例中,有p53蛋白阳性14例、阴性9例;SACC阴性的22例中,有p53蛋白阳性7例、阴性15例。SACC与p53蛋白的表达呈正相关（r=0.505,P〈0.05）。结论宫颈癌组织中SCCA和p53蛋白呈高表达,二者可能共同参与了宫颈癌的发生发展,SCCA可能是促进p53突变的重要因素。     

人乳头瘤病毒致宫颈癌细胞免疫逃逸机制探讨

目的探讨人乳头瘤病毒（HPV）感染与宫颈癌细胞免疫逃逸之间的关系,揭示HPV在宫颈癌发生、发展中的作用。方法以宫颈癌细胞系,宫颈癌组织及其相应的正常宫颈组织为材料,采用实时聚合酶链反应（PCR）检测HPV mRNA表达水平,采用流式细胞仪进行宫颈癌细胞表面C3b沉积检测。结果在30对配对组织中有26对肿瘤组织HPV表达高于配对的癌旁正常组织,C3b的沉积在HPV感染宫颈癌细胞组明显低于无感染组,尤以老年组变化更为明显,同时宫颈癌细胞的迁徙和侵袭能力在HPV感染组明显升高。结论HPV感染的宫颈癌组织通过抑制C3b的沉积而引发免疫逃逸,在宫颈癌的发生、发展中发挥重要作用。     

HPV感染宫颈病变组织中IL-2、IL-4的表达变化及意义

采用原位杂交法检测高危型HPV16／18阳性的宫颈上皮内瘤变（CIN,55例）、宫颈癌（20例）及高危型HPV16／18阳性和阴性的宫颈炎（各15例）组织中的IL-2 mRNA、IL-4 mRNA。结果显示,与高危HPV阴性及阳性宫颈炎组织、高危HPV阳性CINⅠ、CINⅡ组织相比,高危型HPV16／18阳性的CINⅢ和宫颈癌组织中IL-4 mRNA的表达明显上升,IL-2 mRNA则明显下降,P均〈0．01。认为感染高危型HPV16／18的宫颈组织容易发展为CINⅢ甚至宫颈癌,其机制可能是局部细胞因子表达发生变化。     

宫颈癌患者宫颈脱落细胞HPV、病变组织Brn-3a的表达变化及意义

目的观察宫颈癌脱落细胞中人乳头状瘤病毒（HPV）及病变组织中Brn-3a的表达变化,并探讨其临床意义。方法选择经病理确诊的19例宫颈癌、55例CIN（CINⅠ级20例、CINⅡ级16例和CINⅢ级19例）和30例宫颈炎患者,采用基因芯片技术检测其宫颈脱落细胞中的HPV,采用免疫组化SP法检测宫颈病变组织中的Brn-3a。结果宫颈炎、CINⅠ级、CINⅡ级、CINⅢ级和宫颈癌患者宫颈脱落细胞中HPV阳性率分别为43.3%、50.0%、68.8%、84.2%、94.7%,宫颈组织中Brn-3a阳性率分别为6.7%、15.0%、62.5%、78.9%、89.5%。CINⅡ~Ⅲ级及宫颈癌组患者HPV、Brn-3a阳性率均明显高于宫颈炎和CINⅠ患者（P均〈0.01）。宫颈癌患者宫颈脱落细胞HPV及病变组织中Brn-3a的表达呈正相关（r=0.283,P〈0.01）。结论宫颈癌脱落细胞中HPV及病变组织中的Brn-3a高表达,两项指标联合检测有助于宫颈癌前病变的诊断。     

B7-H3 and B7x are highly expressed in human prostate cancer and associated with disease spread and poor outcome

B7-H3 and B7x are recently discovered members of the B7-CD28 family thought to dampen peripheral immune responses via negative costimulation. We evaluated their potential expression in human prostate cancer using a large cohort of patients with 7 years of follow-up. We identified 823 patients with tissue available treated with radical prostatectomy between 1985 and 2003. Immunohistochemistry was performed on tissue microarray sections using anti-B7-H3 and -B7x. The percentage and intensity of immunoreactivity by tumor cells were blindly evaluated by two urological pathologists, and outcome analyses were conducted. Both B7-H3 and B7x were highly expressed; 93% and 99% of tumors had aberrant expression, respectively. The median percentage of tumor cells staining positive was 80% for each molecule. Strong intensity for B7-H3 and B7x was noted in 212 (26%) and 120 (15%) patients, respectively. Patients with strong intensity for B7-H3 and B7x were significantly more likely to have disease spread at time of surgery (P < 0.001 and P = 0.005, respectively). Additionally, patients with strong intensity for B7-H3 and B7x were at significantly increased risk of clinical cancer recurrence (P < 0.001 and P = 0.005) and cancer-specific death (P = 0.004 and P = 0.04, respectively). To our knowledge, we present the largest investigation of B7 family molecules in a human malignancy and a previously undescribed evaluation of B7x in prostate cancer. B7-H3 and B7x are abundantly expressed in prostate cancer and associated with disease spread and poor outcome. Given the proposed immune-inhibitory mechanisms of B7-H3 and B7x, these molecules represent attractive targets for therapeutic manipulation in prostate cancer.     

早期反应基因在宫颈癌中表达及其与人乳头瘤病毒感染的关系

目的探讨早期反应基因(Iex-1)在宫颈癌中表达及与人乳头瘤病毒(HPV)感染的关系。方法病理科留存的宫颈病变组织石蜡包块61份,免疫组化法检测宫颈良性病变、宫颈上皮内瘤变、宫颈癌组织中Iex-1表达,分析Iex-1表达与宫颈癌临床病理特征的关系。培养3种不同HPV感染情况的宫颈癌细胞株C-33A(HPV-)、SIHA(HPV16+)、HELA(HPV18+),实时定量聚合酶链反应(RT-PCR)法和Western印迹法检测各宫颈癌细胞株中Iex-1 mRNA和蛋白表达;使用siRNA靶向沉默SIHA细胞中E6后检测Iex-1 mRNA和蛋白表达,分析其与HPV感染的关系。结果宫颈良性病变、宫颈上皮内瘤变、宫颈癌组织中Iex-1表达阳性率依次下降,差异有统计学意义(P<0.01)。分化程度为G1~G2的宫颈癌患者Iex-1阳性率明显高于分化程度为G3的患者(P<0.05);无宫旁浸润、有淋巴结转移的宫颈癌患者Iex-1阳性率明显高于宫旁浸润、无淋巴结转移的患者(P<0.05)。C-33A细胞株Iex-1 mRNA表达水平明显高于SIHA、HELA细胞株(P<0.01);C-33A细胞株Iex-1蛋白表达水平明显高于SIHA、HELA细胞株(P<0.01)。SIHA细胞株和HELA细胞株Iex-1 mRNA和蛋白表达差异无统计学意义(P>0.05)。干扰组SIHA细胞中E6 mRNA和蛋白表达水平明显低于空白对照组、阴性对照组(P<0.05);干扰组SIHA细胞中Iex-1 mRNA和蛋白表达水平明显高于空白对照组、阴性对照组(P<0.05)。空白对照组与阴性对照组E6 mRNA和蛋白、Iex-1 mRNA和蛋白表达水平差异无统计学意义(P>0.05)。结论Iex-1在宫颈癌组织中低表达,且表达水平与宫颈癌病变程度呈负相关;HPV可能经E6蛋白参与调控的信号通路下调了Iex-1表达,促进宫颈癌发展与转移;首次提出了抑制Iex-1的促凋亡作用可能为HPV诱发宫颈癌的新通路。     

Potential role of soluble B7-H3 in liver immunopathogenesis during chronic HBV infection

Immune-mediated mechanisms have been implicated in liver pathogenesis and subsequent progression in hepatitis B virus (HBV) infection. Costimulatory molecules, the important regulators of immune responses, participate in the regulation of liver pathology in HBV infection. However, the role of B7-H3 (CD276, a new member of B7 family) in this process has not been investigated. In this study, we detected abundant soluble B7-H3 (sB7-H3) in the plasma of patients with chronic HBV infections. The increase of the plasma B7-H3 was associated with the progression of liver cirrhosis and accompanied by decreased expression of B7-H3 on hepatocytes. The identification analysis suggests that the plasma B7-H3 might be derived from the membrane-bound B7-H3 on hepatocytes. A functional study showed that immobilized (4Ig) B7-H3Ig fusion protein could inhibit TCR-induced proliferation and IFN-γ secretion of T cells, which could be partially blocked by soluble B7-H3flag fusion protein. These results suggest that the reduced expression of B7-H3 in the livers might temper the inhibition of T-cell responses mediated by B7-H3 expressed on hepatocytes and thus promote the hepatic inflammation and hepatitis progression in the chronic HBV-infected patients.     

B7-H4 expression in renal cell carcinoma and tumor vasculature: associations with cancer progression and survival

B7-H4 is a recently described B7 family coregulatory ligand that has been implicated as an inhibitor of T cell-mediated immunity. Although expression of B7-H4 is typically limited to lymphoid cells, aberrant B7-H4 expression has also been reported in several human malignancies. To date, associations of B7-H4 with clinical outcomes for cancer patients are lacking. Therefore, we examined B7-H4 expression in fresh-frozen tumor specimens from 259 renal cell carcinoma (RCC) patients treated with nephrectomy between 2000 and 2003 and performed correlative outcome analyses. We report that 153 (59.1%) RCC tumor specimens exhibited B7-H4 staining and that tumor cell B7-H4 expression was associated with adverse clinical and pathologic features, including constitutional symptoms, tumor necrosis, and advanced tumor size, stage, and grade. Patients with tumors expressing B7-H4 were also three times more likely to die from RCC compared with patients lacking B7-H4 (risk ratio = 3.05; 95% confidence interval = 1.51-6.14; P = 0.002). Additionally, 211 (81.5%) specimens exhibited tumor vasculature endothelial B7-H4 expression, whereas only 6.5% of normal adjacent renal tissue vessels exhibited endothelial B7-H4 staining. Based on these findings, we conclude that B7-H4 has the potential to be a useful prognostic marker for patients with RCC. In addition, B7-H4 represents a target for attacking tumor cells as well as tumor neovasculature to facilitate immunotherapeutic treatment of RCC tumors. Last, we demonstrate that patients with RCC tumors expressing both B7-H4 and B7-H1 are at an even greater risk of death from RCC.     

B7-H1 is up-regulated in HIV infection and is a novel surrogate marker of disease progression

The ligation of programmed death-ligand 1 (B7-H1) to T cells results in the preferential production of interleukin 10 (IL-10). We investigated if B7-H1 would be up-regulated in HIV infection, a disease characterized by increased IL-10 production, by measuring B7-H1, B7-1 (CD80), and B7-2 (CD86) expression and mRNA in 36 HIV-infected patients and in 22 healthy controls (HCs). Results showed that (1) B7-H1 expression and mRNA are augmented in cells of HIV patients; (2) increased IL-10 production in these patients is largely induced by B7-H1-expressing CD14(+) cells; (3) an inverse correlation is detected between B7-H1 expression and CD4 counts, whereas the up-regulation of B7-H1 is directly associated with HIV plasma viremia; (4) antiviral therapy results in the parallel down modulation of IL-10 production and B7-H1 expression/synthesis; and (5) B7-H1/CD80 and B7-H1/CD86 mRNA ratios are increased in peripheral blood mononuclear cells (PBMCs) of HIV patients compared with HCs. B7-H1 synthesis and expression are up-regulated in HIV infection, and the degree of dysregulation correlates with the severity of disease. Aberrant antigen presentation by antigen-presenting cells (APCs) that exhibit increased B7-H1 expression and IL-10 production in HIV infection could be responsible for T-lymphocyte unresponsiveness and loss of protective immunity. B7-H1 is a surrogate marker potentially involved in AIDS disease progression.     

Hepatic B7 homolog 1 expression is essential for controlling cold ischemia/reperfusion injury after mouse liver transplantation

Ischemia/reperfusion (I/R) injury remains a key risk factor significantly affecting morbidity and mortality after liver transplantation (LT). B7 homolog 1 (B7-H1), a recently identified member of the B7 family, is known to play important roles in regulating local immune responses. We hypothesized that B7-H1 plays crucial roles during innate immune responses induced by hepatic I/R injury, and using B7-H1 knockout (KO) liver grafts, we tested this hypothesis in the mouse LT model with 24 hours of cold storage. Cold I/R injury in wild type (WT)-to-WT LT enhanced constitutive B7-H1 expression on dendritic cells and sinusoidal endothelial cells and promptly induced B7-H1 on hepatocytes. When B7-H1 KO liver grafts were transplanted into WT recipients, serum alanine aminotransferase (ALT) and graft necrosis levels were significantly higher than those after WT-to-WT LT. Augmented tissue injury in B7-H1 KO grafts was associated with increased frequencies and absolute numbers of graft CD3(+) T cells (particularly CD8(+) T cells). B7-H1 KO grafts had significantly fewer annexin V(+) CD8(+) T cells, and this indicated a failure to delete infiltrating CD8(+) T cells. To evaluate the relative contributions of parenchymal cell and bone marrow-derived cell (BMDC) B7-H1 expression, we generated and transplanted into WT recipients chimeric liver grafts lacking B7-H1 on parenchymal cells or BMDCs. A selective B7-H1 deficiency on parenchymal cells or BMDCs resulted in similar levels of ALT and liver injury, and this suggested that parenchymal cell and BMDC B7-H1 expression was involved in liver damage control. Human livers up-regulated B7-H1 expression after LT. CONCLUSION: The study demonstrates that graft tissue expression of B7-H1 plays a critical role in regulating inflammatory responses during LT-induced hepatic I/R injury, and negative coregulatory signals may have an important function in hepatic innate immune responses.     

Expression and clinical significance of high risk human papillomavirus and invasive gene in cervical carcinoma

Objective:To study the expression of E6 and E7 mRNA in high-risk human papillomavirus(HPV) HPV-18 and the relationship between the expression of invasive gene and cervical carcinoma.Methods:A total of 119 patients with cervical cancer,cervical erosion and cervical HPV infection who were diagnosed in our hospital were selected and randomly divided into two groups:cervical cancer group(n= 58) and non-cancerous group(n= 61).Another 60 patients with uterine leiomyoma were selected as normal control group.Detection of HPV18 E6,E7 mRNA expression and invasion,migration,proliferation inhibition genes,epithelial mesenchymal transition genes and proliferation related protein content.Results:The relative expression of E6 and E7 HPV-18 in cervical cancer group was significant higher than that in non-cancerous group and control group(mRNA)(P0.05).The content of TRAF6 and c-FLIP in invasive cervical cancer group was significantly higher than that in non-cancerous group and control group(P0.05).The mR NA content of CD44v6 and MMP-9 in cervical cancer group was significantly higher than that in non-cancerous group and control group(P0.05).The content of DEC-1,IKK16,MBP-1 in cervical cancer group was significant lower than that in non-cancerous group and control group(P0.05).The mR NA content of beta-catenin and Vimentin in cervical cancer group was significantly lower than that in non cancerous group and control group(P0.05).The proliferation related protein E2F1 of cervical cancer group was significantly lower than that of non-cancerous group and control group,Bmi-1 content was significantly higher than non-cancerous group and control group(P0.05).Conclusions:The expression of the detection of cervical cancer in high-risk human papilloma virus HPV-18 E6 and E7 mRNA,and the invasion,migration,proliferation inhibition gene,epithelial mesenchymal transition and proliferation related gene protein content,HPV expression rate of mR NA increased with the development of cervical cancer,the expression is also enhanced.The expression has a certain correlation between the level and development of cervical cancer.Through the above indicators,the development of cervical cancer monitoring and treatment to provide important clinical guidance.     

B7-H4在上皮性卵巢癌中的表达及临床意义

目的研究B7-H4在原发上皮性卵巢癌（EOC）中的表达情况及与临床生物学特征之间的关系。方法运用免疫组织化学PV-6000二步法，研究B7．H4在70例EOC、20例正常卵巢组织和20冽良性卵巢肿瘤中的表达情况。结果EOC中B7-H4呈胞质和（或）胞膜表达，阳性率为88．57％（62／70），显著高于卵巢良性肿瘤（45％）及正常卵巢组织（0％）中的表达，差异有统计意义（P均〈0．05）。卵巢癌组织中B7-H4表达与组织分化的高低、病理类型及是否淋巴结转移有关。结论B7．H4在EOC中高表达，组织分化越低其表达率越高，淋巴结转移者的表达率增高，B7-H4有望成为卵巢癌新的诊断和预后指标。