胰腺癌中磷酸酶和张力蛋白同源物基因及B7-H1蛋白的表达及其临床意义

Objective To investigate the probable correlation between the expressions of phosphatase and tensin homologue deleted on chromosometen (PTEN) and B7-H1 protein in pancreatic carcinoma and the biological behavior characteristics of tumors. Methods Forty-three patients were recruited who had undergone surgical resection for pancreatic carcinoma between 2002 and 2009. The PTEN and B7-H1 protein expressions in the tissue specimens of these 43 patients and 5 non-pancreatic carcinoma people' s pancreatic tissue specimens were evaluated by immunohistochemistry ELPS technique, and the clinical and pathological features of these specimens and the follow-up information were analyzed. Results PTEN expressions were significantly lower in pancreatic carcinoma tissues than in non-pancreatic carcinoma people' s pancreatic tissues but B7-H1 expressions were significantly higher ( P ＜ 0. 01 ). The expression of PTEN was negatively correlated to that of B7-H1 (r = -0.414 ,P ＜0. 01). PTEN and B7-H1 expressions correlated with the pathological grade and tumor-node-metastasis ( TNM ) stage, peripancreatic invasion, regional lymph node involvement,respectively (P＜0. 05). B7-H1 expressions also significantly correlated with the ages (P＜0. 01). Furthermore, PTEN and B7-H1 expressions showed significant prognostic effects (P＜0.01) and there are correlations existed between combined PTEN/B7-H1 expression and prognostic effects (P ＜0. 05). Conclusion The expression of PTEN and B7-H1 may be significantly correlated to the carcinogenesis,development and prognosis of pancreatic carcinoma.     

胰腺癌中磷酸酶和张力蛋白同源物基因及B7-H1蛋白的表达及其临床意义

Objective To investigate the probable correlation between the expressions of phosphatase and tensin homologue deleted on chromosometen (PTEN) and B7-H1 protein in pancreatic carcinoma and the biological behavior characteristics of tumors. Methods Forty-three patients were recruited who had undergone surgical resection for pancreatic carcinoma between 2002 and 2009. The PTEN and B7-H1 protein expressions in the tissue specimens of these 43 patients and 5 non-pancreatic carcinoma people' s pancreatic tissue specimens were evaluated by immunohistochemistry ELPS technique, and the clinical and pathological features of these specimens and the follow-up information were analyzed. Results PTEN expressions were significantly lower in pancreatic carcinoma tissues than in non-pancreatic carcinoma people' s pancreatic tissues but B7-H1 expressions were significantly higher ( P ＜ 0. 01 ). The expression of PTEN was negatively correlated to that of B7-H1 (r = -0.414 ,P ＜0. 01). PTEN and B7-H1 expressions correlated with the pathological grade and tumor-node-metastasis ( TNM ) stage, peripancreatic invasion, regional lymph node involvement,respectively (P＜0. 05). B7-H1 expressions also significantly correlated with the ages (P＜0. 01). Furthermore, PTEN and B7-H1 expressions showed significant prognostic effects (P＜0.01) and there are correlations existed between combined PTEN/B7-H1 expression and prognostic effects (P ＜0. 05). Conclusion The expression of PTEN and B7-H1 may be significantly correlated to the carcinogenesis,development and prognosis of pancreatic carcinoma.     

前列腺液中B7-H3分子表达对t-PSA灰区内炎性PSA升高的鉴别诊断价值

目的 探讨前列腺液(EPS)中B7-H3分子对血清t-PSA灰区(4～10 ng/ml)内炎性PSA升高患者的鉴别诊断价值.方法 选择2009年12月至2010年4月收治的全部慢性前列腺炎(CP)患者和t-PSA灰区内行前列腺穿刺活检患者共116例,年龄19～80岁,平均40岁.CP 91例,年龄19～49岁,平均31岁.其中慢性细菌性前列腺炎(II型)11例、慢性炎症性非细菌前列腺炎(IIIA型)26例、慢性非炎症性非细菌前列腺炎(IIIB型)54例.t-PSA灰区内接受经直肠超声引导下前列腺穿刺活检患者25例,年龄62～80岁,平均71岁,t-PSA(7.21±2.60)ng/ml.其中穿刺病理结果阳性5例,Gleason评分6分2例、7分2例、8分1例;阴性20例,其中伴炎症细胞浸润11例.采用经直肠按摩法提取EPS.酶联免疫吸附法检测各组EPS B7-H3水平.健康男性对照11例,年龄24～46岁,平均30岁.既往无泌尿系不适症状及手术史.结果 对照组、II型组、IIIA型组、IIIB型组EPS中B7-H3水平依次为(49.81±11.54)、(19.33±13.90)、(17.67±15.76)、(25.14±13.44)ng/ml,穿刺阳性组、阴性不伴炎症组、阴性伴炎症组分别为(26.30±16.32)、(30.23±18.42)、(10.11±5.42)ng/ml.CP各组EPS B7-H3水平均低于对照组,差异有统计学意义(P＜0.01).II型组和IIIA型组间差异无统计学意义(P＞0.05),但均显著低于IIIB型组,差异有统计学意义(P＜0.05).穿刺阴性伴炎症组EPS中B7-H3水平与II型组、IIIA型组比较差异无统计学意义(P＞0.05),但显著低于穿刺阳性组及阴性不伴炎症组,差异有统计学意义(P＜0.05).EPS B7-H3表达检测在t-PSA灰区内诊断炎性PSA升高患者的ROC曲线下面积为0.883(P=0.001),当EPS B7-H3值≤16.24 ng/ml时,诊断敏感性为78.6%,特异性为81.8%.结论 EPS B7-H3表达检测可能成为t-PSA灰区内鉴别诊断炎性PSA升高的新指标,从而减少不必要的前列腺穿刺活检.

Abstract：

Objective To investigate the value of B7-H3 in expressed prostatic secretions (EPS) in differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone (4-10 ng/ml). Methods One hundred and sixteen patients from the ages of 19 to 80 years (mean, 40 years) were stu-died. In the group there were 91 chronic prostatitis (CP) patients (mean age 31 years, 19-49 years), including 11 chronic bacterial prostatitis (type II) patients, 26 inflammatory nonbacterial prostatitis (IIIA) patients and 54 noninflammatory nonbacterial prostatitis (IIIB) patients. Transrectal ultrsound guided prostate biopsy was performed on 25 patients (mean age 71 years, 62-80 years) with t-PSA in gray zone (7.21±2.60 ng/ml). Five had positive results, Gleason score was 6 in two cases, 7 in two cases and 8 in one case. Twenty patients had negative results, of whom 11 patients had inflammatory cell infiltration. EPS was collected by transrectal massage, and Enzyme-linked immunosorbent assays (ELISA) were performed for B7-H3 detection. In addition, 11 normal male controls with a mean age of 30 years (24-46 years) were recruited into the study. Volunteers were excluded if they had a history of genitourinary symptoms or surgery.Results The EPS B7-H3 levels of controls, II, IIIA, IIIB groups were 49.81±11.54, 19.33±13.90, 17.67±15.76, 25.14±13.44 ng/ml, respectively. The levels of EPS B7-H3 in positive biopsy, noninflammatory negative biopsy and inflammatory negative biopsy groups were 26.30±16.32, 30.23±18.42, 10.11±5.42 ng/ml, respectively. The highest levels were found in the control group (P<0.01). Compared to the IIIB, B7-H3 levels in II and IIIA groups were significantly lower (P<0.05). There was no significantly difference between II and IIIA groups (P>0.05). The EPS B7-H3 levels in the inflammatory negative biopsy group were statistically lower than in positive biopsy and noninflammatory biopsy groups (P<0.05). But no significant differences were found among inflammatory negative biopsy, II and IIIA groups (P>0.05). Receiver operating curve (AUC=0.883, P=0.001) utilizing EPS B7-H3 levels≤16.24 ng/ml identified patients with inflammatory elevation of PSA with a sensitivity of 78.6% and a specificity of 81.8% from patients with t-PSA in gray zone. Conclusion The EPS B7-H3 detection provides a new way for differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone resulting in a reduction of unnecessary prostate biopsy.     

Diagnosis of deep vein thrombosis after operation for fracture of the proximal femur: comparative study of ultrasonography and venography

Background We compared investigative methods to study the diagnostic capability of ultrasonography. Subjects were 75 patients with proximal femur fractures who underwent surgery from March 2002 to December 2003. Methods Biochemical assays were carried out on days 3 and 7, and D-dimer levels were investigated as a time series. Ascending lower limb venography and ultrasonography were carried out nearly simultaneously in all cases on day 7. Results The incidence of deep vein thrombosis (DVT)-positive findings on venography was 61.3% (46/75). Venography results demonstrated a diagnostic sensitivity of 95.5% and specificity of 91.4% using a D-dimer cutoff of 1μg/ml on postoperative day 7. By ultrasonography, diagnostic sensitivity for DVT was 78.3%, and specificity was 96.5%. Among all proximal area cases, there were 18 limbs with DVT seen on venography among which were 17 limbs in cases demonstrating DVT at the same site on ultrasonography, a correct diagnosis rate of 94.4%. Conclusions For the diagnosis of a thrombus more proximal than the popliteal area, a location at high risk for occurrence of pulmonary embolism, ultrasonography provided results nearly on par with those obtained by venography. Thus, we believe that ultrasonography allowed a more noninvasive, accurate diagnosis and more rapid treatment in cases where D-dimer values were 1μg/ml or more on postoperative day 7.     

顺铂联合加热对体外血液中肝肿瘤细胞的杀灭作用及对红细胞的影响

Objective To investigate the effectiveness of cis-diamminedichloroplatinum (DDP) combined with hyperthennia in killing liver tumor cells and its influence on erythrocytes in vitro. Methods Cultured liver tumor cells (2 ml) were mixed with erythrocyte suspension (10 ml) and then the mixture was separated into 6 centrifuge tubes with 2 ml in each one. The centrifuge tubes were randomly divided into A-F groups and the experiment was repeated for 30 times. Normal saline 2 ml was added in A and D groups. DDP 2 ml (200 μg/ml) was added in B and E groups. DDP 2 ml (400 μg/ml) was added in C and F groups. The cells were then incubated in warm bath of 37 ℃ for 30 min in A, B and C groups and in warm bath of 42 ℃ for 30 min in the other three groups.After hyperthermic treatment, tumor cells were isolated from erythrocytes using density gradient centrifugation, the inhibition rate of tumor cells was determined by MTT method and the clone formation of tumor cells was checked.The erythrocyte osmotic fragility and content of 2,3-diphosphoglyceric acid in erythrocytes were measured. Results The inhibition rate of tumor cells was gradually increased, while the rate of tumor cell clone formation decreased with the increase in the temperature and DDP concentrations ( P < 0.01) . The rate of tumor cell clone formation was more than 98% and no clone formation was tested in group F. There was no significant difference in the content of 2,3-diphosphoglyceric acid in erythrocytes between before and after hyperthermic treatment in group F ( P >0.05 ) . The rate of hemolysis of erythrocytes was less than 1 % in the 0.68 % sodium chloride solution in group F.Conclusion DDP 200 μg/ml combined with hyperthermic treatment with temperature of 42 ℃ for 30 min can make the liver tumor cells lose the capability of proliferation, however, it exerts slight effect on erythrocyte membrane and no influence on the oxygen-carrying capacity of erythrocytes.     

顺铂联合加热对体外血液中肝肿瘤细胞的杀灭作用及对红细胞的影响

Objective To investigate the effectiveness of cis-diamminedichloroplatinum (DDP) combined with hyperthennia in killing liver tumor cells and its influence on erythrocytes in vitro. Methods Cultured liver tumor cells (2 ml) were mixed with erythrocyte suspension (10 ml) and then the mixture was separated into 6 centrifuge tubes with 2 ml in each one. The centrifuge tubes were randomly divided into A-F groups and the experiment was repeated for 30 times. Normal saline 2 ml was added in A and D groups. DDP 2 ml (200 μg/ml) was added in B and E groups. DDP 2 ml (400 μg/ml) was added in C and F groups. The cells were then incubated in warm bath of 37 ℃ for 30 min in A, B and C groups and in warm bath of 42 ℃ for 30 min in the other three groups.After hyperthermic treatment, tumor cells were isolated from erythrocytes using density gradient centrifugation, the inhibition rate of tumor cells was determined by MTT method and the clone formation of tumor cells was checked.The erythrocyte osmotic fragility and content of 2,3-diphosphoglyceric acid in erythrocytes were measured. Results The inhibition rate of tumor cells was gradually increased, while the rate of tumor cell clone formation decreased with the increase in the temperature and DDP concentrations ( P < 0.01) . The rate of tumor cell clone formation was more than 98% and no clone formation was tested in group F. There was no significant difference in the content of 2,3-diphosphoglyceric acid in erythrocytes between before and after hyperthermic treatment in group F ( P >0.05 ) . The rate of hemolysis of erythrocytes was less than 1 % in the 0.68 % sodium chloride solution in group F.Conclusion DDP 200 μg/ml combined with hyperthermic treatment with temperature of 42 ℃ for 30 min can make the liver tumor cells lose the capability of proliferation, however, it exerts slight effect on erythrocyte membrane and no influence on the oxygen-carrying capacity of erythrocytes.     

大鼠TRESK基因重组腺病毒载体的构建

目的 构建大鼠TRESK基因重组腺病毒载体.方法 从大鼠背根神经节细胞中克隆TRESK全长cDNA,进行PCR鉴定及DNA测序验证,构建以CMV启动子转录调控的pAd/CMV/V5DEST-TRESK.转化DH5α大肠杆菌,挑选阳性重组克隆行PCR鉴定并行DNA测序.将测序正确的质粒经PacⅠ酶切线性化,转染包装293T细胞,包装产生腺病毒,逐孔稀释滴度法测定病毒滴度.结果 从大鼠背根神经节细胞中克隆的TRESK全长cDNA为781 bp,DNA测序验证DNA序列与GeneBank 中收录的大鼠TRESK序列完全一致.以pAD-GFP空载体为对照,pAD/CMV/V5-DEST-TRESK gDNA为模板的PCR扩增,目的 片段781 bp,鉴定结果 与预期相符.腺病毒滴度为1.31×109 TU/ml.结论 本研究成功地构建了大鼠TRESK基因重组腺病毒载体:pAD/CMV/V5-DEST-TRESK.

Abstract：

Objective To construct rat TRESK gene recombinant adenovirus expression vector.Methods TRESK full-length cDNA was cloned from rat dorsal root ganglion cells and confirmed by RT-PCR and sequencing. pAd/CMV/V5-DEST-TRESK was then constructed under the control of CMV-promotor. DH5α colibacillus was translated and the positive recombinants were subsequently identified by PCR and DNA sequencing. 293T cells were cotransfected and packed to produce adenovirus. Results The titer of virus was tested using Hole-by-dilution titer method. The full length of TRESK from rat dorsal root ganglion cells is 781 bp. It was demonstrated that the DNA sequencing was completely consistent with TRESK sequencing of rat recorded in GeneBank. The PCR amplification of the pAd/CMV/V5-DEST-TRESK gDNA was matched with pAD-GFP blank vector as anticipated. The titer of the concentrated virus was 1.31×109 TU/ml. Conclusion Rat TRESK gene recombinant adenovirus vector is constructed successfully.     

胰腺癌中磷酸酶和张力蛋白同源物基因及B7-H1蛋白的表达及其临床意义

目的 观察胰腺癌磷酸酶和张力蛋白同源物基因(PTEN)蛋白和B7-H1蛋白的表达,探讨两者与胰腺癌肿瘤生物学特征的关系.方法 采用免疫组织化学ELPS法检测43例胰腺癌患者的组织标本和作为对照的5例非癌胰腺标本中PTEN和B7-H1蛋白的表达水平,分析其与临床和病理学特征的关系.结果 比较非癌胰腺组织,胰腺癌组织中PTEN蛋白表达显著降低,而B7-H1蛋白表达显著升高(P＜0.01),两者的表达还呈现显著负相关(r=-0.414,P＜0.01);两者的表达水平与肿瘤的分化程度、TNM分期、浆膜侵犯及淋巴结转移有关(P＜0.05),B7-H1表达水平还与年龄显著相关(P＜0.01);两者分别的表达水平与患者的预后均显著相关(P＜0.01),两者的联合表达水平与患者的预后相关(P＜0.05).结论 mN和B7-H1蛋白的表达水平与胰腺癌的发生、发展和预后密切相关.

Abstract：

Objective To investigate the probable correlation between the expressions of phosphatase and tensin homologue deleted on chromosometen (PTEN) and B7-H1 protein in pancreatic carcinoma and the biological behavior characteristics of tumors. Methods Forty-three patients were recruited who had undergone surgical resection for pancreatic carcinoma between 2002 and 2009. The PTEN and B7-H1 protein expressions in the tissue specimens of these 43 patients and 5 non-pancreatic carcinoma people' s pancreatic tissue specimens were evaluated by immunohistochemistry ELPS technique, and the clinical and pathological features of these specimens and the follow-up information were analyzed. Results PTEN expressions were significantly lower in pancreatic carcinoma tissues than in non-pancreatic carcinoma people' s pancreatic tissues but B7-H1 expressions were significantly higher ( P ＜ 0. 01 ). The expression of PTEN was negatively correlated to that of B7-H1 (r = -0.414 ,P ＜0. 01). PTEN and B7-H1 expressions correlated with the pathological grade and tumor-node-metastasis ( TNM ) stage, peripancreatic invasion, regional lymph node involvement,respectively (P＜0. 05). B7-H1 expressions also significantly correlated with the ages (P＜0. 01). Furthermore, PTEN and B7-H1 expressions showed significant prognostic effects (P＜0.01) and there are correlations existed between combined PTEN/B7-H1 expression and prognostic effects (P ＜0. 05). Conclusion The expression of PTEN and B7-H1 may be significantly correlated to the carcinogenesis,development and prognosis of pancreatic carcinoma.     

肾移植患者血清中可溶性B7-H3水平的变化及其临床意义

Objective To detect the serum soluble B7-H3 (sB7-H3) in patients before and after renal transplantation, and to investigate the clinical significance. Methods The serum level of sB7-H3 in 34 patients were determined before and 3, 6, 12 months after renal transplantation. Besides, 11 health adults were elected as controls. These 34 patients were divided into two groups according to HLA and/or MICA antibodies using Luminex before operation. After transplantation, all the patients were divided into two groups according to their conditions; group Ⅰ with rejection; group S with stable renal function and no rejection. Results The serum level of sB7-H3 in all 34 patients before operation was significantly higher than health controls [(27. 10 ± 13. 61) μg/L,n = 34 vs (11.61 ±3.77) μg/L,n = 11 ,P ＜0. 01], and significantly higher in patients with antibodies than in those without before operation [(34. 96 ± 17. 37) (μ/L, n = 11 vs (23. 34 ± 9. 75) (μg/L, n = 23, P ＜ 0. 05]. There was no significant difference in the serum level of sB7-H3 between control group and S group after operation (P ＞0. 05). In group Ⅰ, the serum level of sB7-H3 was increased obviously when rejection occurred [(20. 63 ±4. 28) μg/L,n = 12; (18. 95 ±2.98) μg/L,n=6; (28.36 ±19. 83) μg/L,n = 10] , as compared with group S and control group (P＜0. 01), while at the time without rejection, there was no difference among them (P ＞0. 05). Conclusion Monitoring serum sB7-H3 after renal transplantation would be clinically useful in indicating therapeutic effect and outcome of the patients.     

肾移植患者血清中可溶性B7-H3水平的变化及其临床意义

Objective To detect the serum soluble B7-H3 (sB7-H3) in patients before and after renal transplantation, and to investigate the clinical significance. Methods The serum level of sB7-H3 in 34 patients were determined before and 3, 6, 12 months after renal transplantation. Besides, 11 health adults were elected as controls. These 34 patients were divided into two groups according to HLA and/or MICA antibodies using Luminex before operation. After transplantation, all the patients were divided into two groups according to their conditions; group Ⅰ with rejection; group S with stable renal function and no rejection. Results The serum level of sB7-H3 in all 34 patients before operation was significantly higher than health controls [(27. 10 ± 13. 61) μg/L,n = 34 vs (11.61 ±3.77) μg/L,n = 11 ,P ＜0. 01], and significantly higher in patients with antibodies than in those without before operation [(34. 96 ± 17. 37) (μ/L, n = 11 vs (23. 34 ± 9. 75) (μg/L, n = 23, P ＜ 0. 05]. There was no significant difference in the serum level of sB7-H3 between control group and S group after operation (P ＞0. 05). In group Ⅰ, the serum level of sB7-H3 was increased obviously when rejection occurred [(20. 63 ±4. 28) μg/L,n = 12; (18. 95 ±2.98) μg/L,n=6; (28.36 ±19. 83) μg/L,n = 10] , as compared with group S and control group (P＜0. 01), while at the time without rejection, there was no difference among them (P ＞0. 05). Conclusion Monitoring serum sB7-H3 after renal transplantation would be clinically useful in indicating therapeutic effect and outcome of the patients.     

Evaluation of CMV viral load using TaqMan CMV quantitative PCR and comparison with CMV antigenemia in heart and lung transplant recipients.

BACKGROUND: Quantitative assessment of cytomegalovirus (CMV) infection using the antigenemia test has been used to monitor CMV infection in heart and lung transplant patients enabling a preemptive treatment strategy. However, the method is labour intensive, samples have to be processed within a few hours and requires skilled interpretation. A comparative prospective evaluation of a real-time TaqMan CMV quantitative PCR (QPCR) with the CMV antigenemia was undertaken. METHODS: A real-time quantitative TaqMan CMV PCR from EDTA bloods was developed. In this study 25 heart transplant and single-lung transplant patients were monitored posttransplantation by antigenemia and TaqMan CMV QPCR. CMV DNA extracted from EDTA blood was amplified by TaqMan QPCR using primers and probe designed from the CMV glycoprotein B (gB) gene. Quantification of the genome copies is extrapolated from a standard curve generated from amplification of quantified standards. RESULTS: Antigenaemia levels and TaqMan CMV QPCR genome copies showed a linear correlation between the two assays (R=0.843, P=0.001). A clinically significant threshold of 50 CMV pp65 antigen positive polymorphonuclear leucocytes (PMNLs) per 200 000 cells previously reported was used to extrapolate an equivalent value of 40 000 (log 4.6) genome copies per ml of blood for the TaqMan CMV QPCR. CONCLUSIONS: The TaqMan system enables a rapid high-throughput of samples. The TaqMan CMV QPCR can be used as an accurate and robust alternative to the antigenemia test to predict CMV disease and to monitor effectiveness of treatment.     

转录因子T-bet与GATA结合蛋白3的比值在大鼠肺移植急性排斥反应中的mRNA表达及其意义

目的 探讨T-bet(T-box expression in T cells)和GATA结合蛋白3(GATA-3)在大鼠肺移植急性排斥反应中的mRNA表达及其意义.方法 建立同种异体大鼠肺移植模型,实验分为正常对照组(n=5只)、同种异体移植3 d组(n=4只)和5 d组(n=6只),用实时荧光定量逆转录-聚合酶链反应(RT-PCR)法,检测移植后肺组织中T-bet和GATA-3的mRNA表达.结果 实时荧光定量RT-PCR检测T-bet和GATA-3的扩增效率为78%～85%,批内和批间实验循环阈值(Ct)变化均＜0.15.与止常对照和右侧非移植肺比较,同种异体移植术后3 d组,T-bet和GATA-3无显著性改变,但其比值明显高于右侧肺;同种异体移植术后5 d组,T-bet表达增高(9.31拷贝/106 18 S拷贝),但差异无统计学意义(P＞0.05),GATA-3表达下降(0.88拷贝/105 18 S拷贝),T-bet/GATA3的比值增高(1.12),差异均有统计学意义(P＜0.05).结论 实时荧光定量RT-PCR检测T-bet和GATA-3结果准确可靠.在急性排斥反应中,T-bet表达升高,GATA-3表达降低,其中GATA-3的表达起主导作用.T-bet/GATA-3的比值比单独检测T-bet或GATA-3更能客观地评价大鼠肺移植急性排斥反应.     

Plasmacytoid Dendritic Cell Precursors Induce Allogeneic T-Cell Hyporesponsiveness and Prolong Heart Graft Survival

Dendritic cell (DC) precursors were propagated from C57BL/10 (B10; H2b) mouse bone marrow in fms-like tyrosine kinase 3 ligand. Cosignaling molecule (B7-1/B7-2 and B7-H1) expression and stimulatory capacity of precursor (pre)-plasmacytoid (p)DC (CD11c+B220+CD11b-CD19-) and classic myeloid DC (MDC) for allogeneic (C3H; H2k) T cells were compared. Unstimulated pre-pDC exhibited very low levels of surface MHC class II and classic costimulatory molecules (B7-1/B7-2), whereas a minor population expressed B7-H1 at levels higher than on MDC. The pre-pDC were ineffective T-cell stimulators and induced nonspecific hyporesponsiveness to rechallenge with donor alloantigens in vitro and in vivo. Following stimulation with CpG-oligonucleotide (CpG-ODN), B7 molecule expression was upregulated on pre-pDC, however the ratio between coinhibitory (B7-H1) and costimulatory (B7-1/B7-2) signals was much higher (five- to six-fold) on pre-pDC than MDC. Blockade of B7-H1 expression on pDC increased their T-cell allostimulatory capacity significantly. A single preoperative infusion of C3H hosts with pre-pDC prolonged B10 heart graft survival significantly but nonspecifically compared with untreated mice (median survival times 22 vs. 9 days, respectively). Thus, pre-pDC of donor origin have potential to regulate T-cell responses to alloantigens and can prolong organ graft survival.     

协同刺激分子B7-H4影响细胞因子诱导的杀伤细胞治疗胃癌预后的多因素COX模型分析

目的 探讨协同刺激分子B7-H4的表达与胃癌的关系,并分析B7-H4在胃癌组织中的不同表达水平对细胞因子诱导的杀伤细胞(CIK)治疗胃癌患者的复发和死亡的风险.方法 对156例胃癌病例(1998年1月1日至2009年12月31日),采用免疫组织化学染色及IHS评估法确定B7-H4的表达状态,使用回顾性队列研究方法调查每位病例的人口学、临床资料及复发时间和死亡时间.应用COX多因素模型分析B7-H4高表达影响胃癌复发和死亡的风险.结果 胃癌组织B7-H4高表达组与B7-H4低表达组比较,患者中位无瘤生存时间(月)及95%CI分别为16(11.9～20.1)和47(22.4～71.6),中位生存时间(月)及95%CI分别为25(19.0～31.1)和43(35.2～50.8).在化学治疗组中,B7-H4低表达患者的中位生存时间明显高于B7-H4高表达患者(36比16),差异有统计学意义(x2=14.14,P<0.01).在化疗联合CIK治疗组中,B7-H4低表达患者的中位生存时间有高于B7-H4高表达患者的趋势(55比34),但差异无统计学意义(x2=1.78,P>0.05).在化学治疗组中,B7-H4低表达患者中位无瘤生存时间高于B7-H4高表达(33比11),差异有统计学意义(x2=18.956,P<0.01).在化疗联合CIK细胞治疗组中,B7-H4低表达患者中位无瘤生存时间显著高于高表达组(90比18),差异有统计学意义(x2=3.842,P<0.05).在调整了性别、年龄等混杂因素后,与B7-H4低表达比较,B7-H4高表达组病例的复发和死亡风险明显增加(RR=2.30,95%CI=1.41～3.75;RR=1.90,95%CI=1.20～3.01).结论 B7-H4高表达可能是提高胃癌复发和死亡风险的独立危险因素.采用CIK细胞回输治疗可控制和干预B7-H4的表达并延长胃癌患者的中位生存时间,胃癌B7-H4低表达可作为CIK细胞治疗的纳入标准.     

B7-H1在直肠癌与镶嵌血管中的表达及意义

目的 探讨共刺激配体B7-H1在直肠癌和镶嵌血管中的表达及临床意义.方法 应用免疫组织化学方法 观察50例直肠癌患者癌组织和镶嵌血管中的B7-H1表达,探讨B7-H1表达和直肠癌的病理学特征及预后间的关系.结果 随访期13例患者死亡,37例存活患者的中位随访期为3.4年(0～5年);64%的标本显示蛋B7-H1阳性染色,中位染色评分为2.5(0～4);B7-H1阳性表达与临床和病理特征呈逆相关(P＜0.05).15例(30%)的肿瘤患者同时B7-H1和镶嵌血管表达阴性,3例(6%)B7-H1阴性而镶嵌血管阳性,13(26%)B7-H1阳性而镶嵌血管阴性,19例(38%)B7-H1和镶嵌血管同时阳性表达.B7-H1阳性的肿瘤患者较N7-H1阴性的患者更可能镶嵌血管表达阳性(59.3%比11.1%;P＜0.01).双阳性组的生存率明显低于阴性或单阳性组(P＜0.01).21例表达镶嵌血管的直肠癌标本中有8例显示镶嵌血管内的内皮细胞或肿瘤细胞存在B7-H1的表达.结论 在直肠癌及镶嵌血管中阳性表达的B7-H1可作为直肠癌患者的预后标志.B7-H1参与直肠癌及镶嵌血管的免疫逃逸.     

肾移植受者BK病毒相关性肾病的临床诊断

目的 探讨肾移植受者BK病毒相关性肾病(BKVAN)的临床诊断方法 .方法 分别于肾移植术后第1、3、6、9、12个月收集90例肾移植受者血、尿标本,进行尿沉渣Decoy细胞计数与BK病毒(BKV)DNA含量的检测.应用免疫组织化学技术检测移植肾组织中的SV40-T蛋白,移植肾组织及确诊为BKVAN患者的尿沉渣中脱落的肾小管上皮细胞进行普通透射电镜观察.结果 90例肾移植受者1年内尿液Decoy细胞、BKV DNA及血浆BKV DNA阳性率分别为:42.2%(38/90)、45.6%(41/90)和22.2%(20/90);阳性者中位数水平分别为8个/10 HPF、2.60×10~5拷贝/ml和9.65×10~3拷贝/ml.确诊BKVAN 5例.结论 肾移植术后定期规律监测有关BKV活动的指标非常必要.确诊BKVAN需依靠普通病理染色与免疫组织化学染色或普通透射电子显微镜并结合临床表现.     

Overexpression of B7-H3 in CD133 colorectal cancer cells is associated with cancer progression and survival in human patients

Cancer stem–like cells are enriched in CD133-positive (CD133⁺) colorectal cancer (CRC) cells. To date, the biological significance of CD133 expression in cancer stem–like cells is still unknown. B7-H3, a costimulatory molecule, plays a pivotal role in tumor immune escape by inhibiting the functions of T cells. To identify a new marker to predict the tumor grade of CRC, we analyzed the expression of B7-H3 and CD133 in colorectal tumor samples, and their clinical significance was determined. By using a series of techniques including pathologic tissue microarray technology, immunohistochemistry, and immunofluorescent staining, we found B7-H3 was expressed in 56.73% of the CRC cases (59/104) sampled; CD133 was detected in 26.92% of the CRC cases (28/104) sampled. Further analysis indicated that 22 of these CD133⁺ samples expressed B7-H3. We also found coexpression of CD133 and B7-H3 in tumor tissue samples (r = 0.321, P < 0.01). Moreover, in contrast to individual CD133 or B7-H3 expression, the coexpression of B7-H3 and CD133 was evidently associated with the depth of tumor invasion, lymphatic metastasis, distant metastasis, and Dukes' stage, suggesting it is a valuable biomarker for the progression of CRC. Indeed, the patients with coexpression of B7-H3 and CD133 had a poorer survival than the other patients (P < 0.05). In summary, our results reveal that B7-H3 was aberrantly expressed in CD133⁺ CRC cells, and the expression level was closely associated with tumor progression.     

共刺激分子B7-1、B7-2在大鼠热缺血再灌注肝脏表达意义的研究

目的:探讨共刺激分子B71和B72在大鼠肝脏热缺血伤再灌注损伤模型中的表达情况及其免疫学意义。方法:将雄性Wistar大鼠随机分为3组:A组(缺血30min再灌注24h)、B组(缺血60min再灌注24h)、C组(假手术组)。模型制备参考Ohmorid的方法。分别取模型之肝左叶,采用实时反转录聚合酶链反应(PTPCR)检测B71和B72在3组肝左叶组织中mRNA表达情况。结果:B71和B72mRNA在C组以极低水平表达,而在A组、B组表达却明显增多(P<0.01),且B组高于A组(P<0.05)。结论:热缺血再灌注之肝脏通过上调B71和B72的表达增加了肝脏的免疫原性。