目的 探讨乙型肝炎病毒（HBV）表面抗原（HBsAg）阳性肝硬化患者血清中HBV前S1抗原(前S1抗原)、HBV e抗原（HBeAg）及HBV核酸定量检测（HBV DNA）相关性。 方法 2008年7月－2011年5月对97例HBsAg阳性肝硬化住院患者和50份HBsAg阴性的健康体检者血清进行前S1抗原、HBV血清标志物检测及实时荧光定量PCR检测HBV DNA结果进行分析。 结果 97份HBsAg阳性肝硬化患者血清中，前S1抗原、HBeAg及HBV DNA阳性率分别为53.6%（52/97）、22.7%（22/97）及61.8%（60/97）。22例HBeAg阳性血清中，前S1抗原阳性18例（81.8%）， HBV DNA阳性20例（90.9%)。75例HBeAg阴性血清中，前S1抗原阳性34例（45.3%），HBV DNA阳性40例（53.3%），两者的前S1抗原与HBV DNA结果间都具有很好的相关性。HBV DNA含量与前S1抗原及HBeAg阳性结果显示：HBsAg阳性的肝硬化患者血清中HBV DNA阴性率为38.1%（含量＜103 copies/mL），而阳性检出率HBV DNA含量主要集中在103～105 copies/mL，占81.7%（49/60），HBV DNA含量＞105 copies/mL占18.3%（11/60）。 结论 HBsAg阳性的肝硬化患者血清中主要以HBV非HBeAg阳性血清学模式为主，HBV DNA阳性检出率的含量主要集中在103～105 copies/mL。前S1抗原在HBeAg阳性血清中与其含有HBsAg病毒及HBeAg阳性患者具有很好的相关性，而在HBeAg阴性血清中存在着差异。Objective To study the correlation among Pre-S1 antigen, HBeAg and HBV DNA results in patients with HBsAg-positive liver cirrhosis. Methods We retrospectively analyzed the serum pre-S1-antigen, HBV serum markers and real-time quantitative PCR HBV DNA results in 97 patients with HBsAg-positive liver cirrhosis and 50 HBsAg-negative healthy volunteers in our hospital from July 2008 to May 2011. Results Among the 97 samples of HBsAg-positive liver cirrhosis patients’ serum, the positive rates of Pre-S1 antigen, HBeAg and HBV DNA were 53.6% (52/97), 22.7% (22/97) and 61.8% (60/97), respectively. In the 22 samples of HBeAg-positive serum, the number of positive pre-S1 antigen and HBV DNA was 18 (81.8%) and 20, respectively. In the 75 samples of negative HBeAg serum, the number of positive pre-S1 antigen and HBV DNA was 34 (45.3%) and 40 (53.3%) respectively. The pre-S1 antigen was correlated well with HBV DNA results in both the two groups. HBV DNA level, pre-S1 antigen and HBeAg-positive results showed that the serum HBV DNA negative rate of HBsAg-positive patients with cirrhosis was 38.1% (＜103 copies/mL), while the positive rate of HBV DNA level was mainly concentrated at 103~105 copies/mL, accounting for 81.7% (49/60), and HBV DNA level over 105 copies/mLaccounted for only 18.3% (11/60). Conclusions HBsAg-positive patients with cirrhosis mainly have a serum non-HBeAg-positive HBV serology pattern, and HBV DNA positive rate of the content is mainly concentrated at 103~105 copies/mL. There is a good correlation between pre-S1 antigen in HBeAg-positive serum and patients with HBsAg virus or positive HBeAg, while for Pre-S1 antigen in HBeAg-negative serum, it is quite different.
【摘要】 目的 探讨葡萄糖转运蛋白Ⅰ型（glucose transporter 1，GLUT1）和肿瘤增殖细胞核抗原Ki-67在卵巢上皮性肿瘤组织中的表达及其临床意义。 方法 收集2000年1月-2008年6月不同卵巢上皮性肿瘤病变患者119例的组织标本，采用免疫组织化学SP二步法检测肿瘤组织中GLUT1和Ki-67的表达情况。 结果 卵巢交界性、恶性上皮性肿瘤灶性或广泛高表达GLUT1和Ki-67，其表达强度有差异。卵巢良性上皮性肿瘤不表达GLUT1和Ki-67。在卵巢癌中GLUT1及Ki-67的表达强度与病理分级、临床分期、预后有关。GLUT1表达强度与病理分型无关，Ki-67表达强度与病理分型有关。 结论 卵巢上皮性肿瘤组织中GLUT1和Ki-67的表达具有相关性，其表达强度与肿瘤的良恶性质和增殖状态有关，二者同时检测可以全面了解卵巢上皮性肿瘤的性质、卵巢癌恶性程度和生物学行为，对于判断肿瘤的性质和预后有一定价值。【Abstract】 Objective To investigate the expression and clinical significance of glucose transporter-1 (GLUT1) and tumor proliferating karyon antigen Ki-67 in epithelial ovarian tumor tissue. Methods Immunohistochemistry SP method was used to detect the expression of GLUT1 and Ki-67 protein in epithelial ovarian tumor tissues from 119 patients diagnosed in our hospital from January 2000 to June 2008. Results The expressions of GLUT1 and Ki-67 had local or abroad higher expressions in the borderline and malignant epithelial ovarian tumor, and the expressive intensity was different. In benign tumors, the expression was negative. The expressive intensity of GLUT1 and Ki-67 had correlation with the grade, stage, and prognosis in malignant tumors. The expressive intensity of GLUT1 had no correlation with the type of malignant tumors, while Ki-67 related to the pathological types. Conclusion The expressions of GLUT1 and Ki-67 have relativity. The expressive intensity of GLUT1 and Ki-67 relates to the character and proliferation of epithelial ovarian tumors. The combined detection GLUT1 and Ki-67 is helpful to know the character of epithelial ovarian tumors, the malignant degree and biologic behavior of ovarian carcinoma, which is useful in estimating the character and prognosis of epithelial ovarian tumors.
目的：建立能表达乙肝病毒（hepatitis B virus,HBV）preS2S抗原蛋白的荷瘤小鼠模型，为研究HBV核酸疫苗的体内CTL应答及免疫治疗作用提供简便易行的研究模型。 方法：以免疫印迹法验证SP2/0-S2S细胞中有HBV preS2S抗原的稳定表达，将SP2/0-S2S细胞种植到BALB/c小鼠胁部皮下（荷瘤），观察能否生长成瘤，以及成瘤的时间、肿瘤大小和荷瘤后小鼠的生存时间；以免疫组织化学法检测小鼠肿瘤组织HBV preS2S抗原的表达。以不表达preS2S抗原蛋白的SP2/0-CMV细胞荷瘤小鼠作为阴性对照。结果：荷瘤后3天～1周，SP2/0-S2S细胞可在小鼠皮下形成实体肿瘤，成瘤率为100%，肿瘤细胞中有preS2S抗原表达，荷瘤后小鼠的平均生存时间为16±1天；与不表达preS2S抗原蛋白的SP2/0-CMV细胞荷瘤小鼠相比，成瘤率、成瘤时间、肿瘤大小及生存时间差异。结论：建立了能表达HBV preS2S抗原蛋白的荷瘤小鼠模型，可用于HBV核酸疫苗的体内CTL应答及免疫治疗作用的实验研究。同时也建立了不表达preS2S抗原蛋白的荷瘤小鼠模型，可用作阴性对照。
Objective To explore the effect of cytotoxic T lymphocyte-associated antigen 4 (CTLA4-Ig) fusion protein on the function of orthotopic liver allograft. Methods Orthotopic liver allograft models of rhesus monkeys were established in this study. The survival time, liver function and morphologic changes of graft were observed, respectively. The levels of IL-2 and IL-10 were detected by ELISA. Apoptosis was monitored by TUNEL.Results The average survival of control group was 6.57 d, while the average survival of CTLA4-Ig group was 14.92 d, which was statistically prolonged (Plt;0.05). Serum ALT level was highly increased, and Alb level decreased obviously in control group. While the levels of ALT and Alb kept in normal in CTLA4-Ig group. After day 3-7, the expressions of IL-2 were highly expressed in control group, while the expressions of IL-10 in CTLA4-Ig group were higher than those of control group. The severity of rejection reaction after day 3 was weaker in CTLA4-Ig group than that of control group by histological assessment. The apoptosis index after day 3 in the liver cells was highly increased in control group as compared with the CTLA4-Ig group. Conclusions CTLA4-Ig fusion protein therapy can induce immunotolerance and prolong the survival of recipients. The increasing of cytokines IL-2 or the decreasing of cytokines IL-10 may be one of the laboratory indexes in monitoring immunotolerance of transplantation.
Objective To study the expression of proliferating cell nuclear antigen (PCNA) in the occurrence and development of hepatocellular carcinoma. Methods Sixty SD rats were randomly divided into control group and experimental group. 3′MeDAB was administrated into rats to establish the experimental model of hepatocarcinoma. The expressions of PCNA of different phases were detected by immunohistochemistry and the liver pathologic changes were observed by optical microscope. Results The process of canceration was divided into three stages: inflammation, proliferative fibrosis and hepatic carcinoma. The expression of PCNA firstly presented in the oval cells that located in the portal area at the stage of inflammation, and a part of PCNA were hyperexpressed in the portal area. The expression rate of PCNA in the middle phase of inflammatory stage was higher than that of any other phases but declined later. Yet, when it came to the stage of hepatic carcinoma, the rate increased again. Conclusion Under the experimental circumstance when liver cancer is caused by the carcinogenic agent, PCNA may be firstly expressed in the oval cells, and the dynamic expression of PCNA may be an indicator for the early diagnosis of hepatocarcinogenesis.
Objective To study the effect of HBeAg on recurrence and survival after radical resection of small (≤3 cm) hepatocellular carcinoma (HCC). Methods Two hundreds and twenty-three HCC patients undergone radical resection from 1999 to 2000 were divided into two groups according to serum HBeAg status， HBeAg positive group (n=73) and HBeAg negative group (n=150). The patients’ factors, operative factors and tumorous facors were studied retrospectively between the two groups. And risk factors of overall survival (OS) and disease-free survival (DFS) were analyzed. Results There were no significant differences in operative and tumorous factors between the two groups, but the HBeAg positive group were younger with more severe cirrhosis (P=0.004, P=0.008). The OS and DFS were significantly different between the HBeAg positive group and HBeAg negative group. The 1-, 3- and 5-year OS were 91.5%, 76.8%, 60.1% and 95.2%, 85.3%, 73.2%, respectively (P=0.053)； and the 1-, 3- and 5-year DFS were 73.3%, 53.7%, 40.3% and 86.6%, 65.5%, 54.5%, respectively (P=0.002). Multivariate analysis revealed that age ＞50 years, HBeAg positive and macronodular cirrhosis were significantly related to OS, and HBeAg positive, multiple tumor nodulars were significantly related to DFS. Positive serum HBeAg status was an independent risk factor for both OS and DFS. Conclusion Positive serum HBeAg is closely related to early recurrence and survival after radical resection of patients with small HCC.
【Abstract】Objective To investigate the expression of PCNA in gastric cancer and its relationship with telomerase activity of peritoneal washings and peritoneal dissemination, and to compare the efficacy of telomerase activity and cytology of peritoneal washings for prediction of peritoneal metastasis of gastric cancer. MethodsTelomeric repeated amplification protocol (TRAP)enzymelinked immunosorbent assay (ELISA) was performed to measure the telomerase activity of peritoneal washings collected from 60 patients with gastric cancer. Exfoliate cytologic analysis of the corresponding samples was used for comparison.Expression of PCNA was measured with immunohistochemical staining.Their relationship with clinicopathologic features were evaluated. ResultsThe positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was 41.7%,which well related to serosal invasion, histology types, depth of infiltration and peritoneal metastasis of gastric cancer. The positive rate of telomerase activity increased with the increased depth of infiltration and serosal involvement areas (P＜0.05).The positive rate of exfoliative cytology was 25.0%, which was obviously high in the group with macroscopic peritoneal metastasis (the group of P1-3). The positive rate of exfoliative cytology also increased with the increased depth of infiltration and serosal involvement areas (P＜0.05). Although the positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was not significantly higher than that of exfoliative cytology in general, it was significantly higher than that of exfoliative cytology in the group of pT4, P1-3 and undifferentiated type.The PCNA proliferation index (PI) of positive telomerase activity group was significantly higher than that of negative. The PCNA PI was significantly higher in the group of P1-3 and serosal invasion thanthat of P0 and without serosal invasion. ConclusionTo detect telomerase activity in peritoneal washings and to detect tumor cells by cytologic method are useful to predict subclinical metastasis to the peritoneum in patients with gastric cancer,but telomerase activity is more sensitive than the other one.Telomerase activity is well related to proliferating activity of gastric cancer,which was the very important reason of peritoneal metastasis and serosal invasion.
【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.