Objective
To investigate the correlation of expression of Fas/Fas ligand (FasL) and apoptosis in retinoblastoma (RB)．
Methods
The expression and distribution of Fas/FasL were detected by using immunohistochemical staining in 32 cases of RB． Light microsc opy (32 cases), electron microscopy (4 cases) and TdT mediated biotin-d UTP nick-end labeling (TUNEL) (12 cases) were used to study apoptosis in RB．
Results
Apoptotic RB cells mostly located at RB regress area． Chromatin margination and apoptotic bodies were found in RB． TUNEL posi tive labeling cells especially located in tumor regress area． Positive immunola beling for Fas and FasL was found in all RB specimens． There was a highly signi ficant and positive correlation between the expression of Fas/FasL and apoptotic indices (AI) (Plt;0.01 or 0.001)．
Conclusion
The results suggest that apoptotic cell death is prevalent in RB and it may be one type of the most dominant cell death． Fas system may play an important role in oncogenesis and progression of RB, and the up-regulation of Fas system expression might induce RB cell apoptosis．
(Chin J Ocul Fundus Dis, 2001,17:21-23)
ObjectiveTo investigate the expression and distribution of CD15s antigen in breast cancer and its relationship with carcinogenesis, progression and metastatic proclivity. MethodsCatalyzed signal amplification(CSA) immunohistochemical technique was used to detect the expression of CD15s antigen in breast cancer and in adjacent normal mucosa. Immunoelectromicroscopic ultrastructural localization of CD15s antigen labelled by colloidal gold was also bserved.ResultsThe positive rate of CD15s antigen expression in primary breast cancer was 79.8%(75/94). In adjacent normal mucosa (n=10) CD15s antigen showed weaker staining. The positive rate of CD15s antigen expression in grade Ⅱ-Ⅲ (87.3%) was notably higher than that in grade Ⅰ (69.2%, P＜0.05). In patients with lymph node metastasis, the positive rate of CD15s antigen expression was 90.2%, which was significantly higher than 67.4% in nodes with no metastasis (P＜0.05). CD15s antigen immunoreactivity was mainly localized in the border membrane of cytoplasm, endoplasmic reticulum, golgi complex and surrounding nuclear membrane in tumor tissue, and in the border membrane of cytoplasm in adjacent normal tissue. Conclusion CD15s antigen is a practical parameter for evaluating the degree of malignancy and lymphatic metastatic proclivity of breast cancer. It can provide a new pathway to investigate the carcinogenesis and progression of breast cancer.
Objective To establish a eukaryotic cell line that can express soluble human leucocyte antigen G1(sHLA-G1) stably. Methods The recombinant plasmid pcDNA3-sHLA-G1 is transfected by a novel nonviral, electroporation-based gene transfer method termed nucleofection into the host cell lymphoblastoid cell line (LCL)721.221 which does not express any HLA-classical I molecules. After selection by G418, the cell line stably expressingsHLA-G1 is identified by RTPCR and Dot-ELISA with HLA-G1 specific monoclonal antibody MEM-G/9. Results The efficiency of transfection for LCL721.221 is about 14% by nucleofection. The specific band forsHLA-G1 was found by RT-PCR assay from the transfections and the protein ofsHLA-G1 in the supernatant of the transfections was detected by Dot-ELISA assay. Both confirmed that the eukaryotic cell line expressingsHLA-G1 has been established successfully at genic and proteinic levels. Conclusion In this study, the eukaryotic cell line expressingsHLA-G1 have been established successfully by nucleofection.
ObjectiveTo summarize the results of testing and analysis of antigen and antibody for diseases under the frame of children's immunization program, in order to know the effects of prevention and control of such diseases in this area.
MethodsA total of 150 children from each of the 5 communities or administrative villages in Yongning District of Nanning City were selected for our survey between January and December 2012. The 150 children were composed of 30 children (residents, 1-6 years old, 5 children from each different age group) randomly selected from each of the four directions (east, south, west and north) and the mid-area of each community or village. The serum samples were collected to analyze the existence of poliomyelitis antibody, measles antibody, hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), and diphtheria antibody.
ResultsAmong the 150 school-age children, antibody immune qualified rate was high for hepatitis B (HBV) antigen, in which the HBsAg immune qualified rate was 99.3%, and HBsAb immune qualified rate was 88.0%, showing no significant difference between boys and girls (P>0.05). All poliomyelitis Ⅰ, Ⅱ, and Ⅲ antibody positive rates reached 100.0%. Measles antibody test results were also satisfying for each age group, among whom the 2 and 3 year-olds reached a positive rate of the highest, 100%, and the 1, 4, 5, and 6 year-old children had a measles antibody positive rate of 96.0%, 84.0%, 88.0%, and 96.0%, respectively. The positive rate for diphtheria antibody was 100%.
ConclusionThe antibody and antigen detection and analysis results for the children's immune program targeted diseases are generally satisfying in this area. Especially, the prevention of poliomyelitis and diphtheria is the best. However, prevention of HBV and measles is not as good. Therefore, tracking immunization coverage, promoting public awareness on immune planning, actively participating in the vaccination of children should be enhanced for further disease prevention.
【Abstract】Objective To investigate the expression of the mRNA of cancer-testis antigen 9 (CT9) gene in hepatocellular carcinoma. Methods The expression of CT9 mRNA was detected through RT-PCR in HCC tissues and their adjacent non-HCC tissues from 45 HCC patients. From CT9 RT-PCR positive products, 3 samples were selected randomly and were sequenced. ResultsCT9 mRNA was detectable in 51.1%(23/45) of HCC samples, and no expression of CT9 mRNA was detected in the adjacent non-HCC tissues. In addition, the RTPCR products were proved to be CT9 cDNA by DNA sequencing. No relationship was found between the expression of CT9 mRNA and clinical factors such as age, sex, tumor size, degree of tumor differentiation, serum αfetoprotein level and infection of hepatitis B virus or hepatitis C virus （Pgt;0.05）. ConclusionCT9 mRNA is expressed with high percentage and specificity in hepatocellular carcinomas. The CT9 gene product is a potential target for antigenspecific immunotherapy of HCC.
目的 探討乙型肝炎病毒（HBV）表面抗原（HBsAg）陽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核酸定量檢測（HBV DNA）相關性。 方法 2008年7月－2011年5月對97例HBsAg陽性肝硬化住院患者和50份HBsAg陰性的健康體檢者血清進行前S1抗原、HBV血清標志物檢測及實時熒光定量PCR檢測HBV DNA結果進行分析。 結果 97份HBsAg陽性肝硬化患者血清中，前S1抗原、HBeAg及HBV DNA陽性率分別為53.6%（52/97）、22.7%（22/97）及61.8%（60/97）。22例HBeAg陽性血清中，前S1抗原陽性18例（81.8%）， HBV DNA陽性20例（90.9%)。75例HBeAg陰性血清中，前S1抗原陽性34例（45.3%），HBV DNA陽性40例（53.3%），兩者的前S1抗原與HBV DNA結果間都具有很好的相關性。HBV DNA含量與前S1抗原及HBeAg陽性結果顯示：HBsAg陽性的肝硬化患者血清中HBV DNA陰性率為38.1%（含量＜103 copies/mL），而陽性檢出率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）。 結論 HBsAg陽性的肝硬化患者血清中主要以HBV非HBeAg陽性血清學模式為主，HBV DNA陽性檢出率的含量主要集中在103～105 copies/mL。前S1抗原在HBeAg陽性血清中與其含有HBsAg病毒及HBeAg陽性患者具有很好的相關性，而在HBeAg陰性血清中存在著差異。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.
Objective
To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo.
Methods
The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed.
Results
① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05).
Conclusion
CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.
Objective To investigate the relationship between human leukocyte antigen(HLA)-B51 and Behcet′s disease (BD) with uveities. Methods HLA-A and HLA-B antigen of 27 pateints with BD and 30 healthy persons were detected by microly mphocyte toxicity asssay. HLA-B51 allele (HLA-B5101-B5107) in BD patients with positive HLA-B51 antigen and the controls was detected by polymerase chain reaction-sequence specific primer(PCR-SSP). Results The positive rate of HLA-B51 was 63% and 16.7% in BD patients and the controls, respectively (χ2=12.9, P＜0.001, Pc＜0.05,RR=8.5). Uveities was found in 11 out of 27 BD patients with uveitis. No relativity was found between HLA-B51 and uveitis in BD patients(RR=2.07,χ2=0.759,P＞0.25),and weak association was found between HLA=B5101 and uveitis (RR=2.67, χ2=1.395, P＞0.10). Conclusions HLA-B51 might be a susceptible gene for BD, and there was a weak association between HLA-B51(HLA-B5101) and BD patients with uveitis.(Chin J Ocul Fundus Dis，2004，20：203-205)
Objective To observe the expression of Ezrin protein in primary carcinoma of gallbladder tissue and the levels of CEA and CA19-9 in serum of patients with primary carcinoma of gallbladder, and to explore the relationship between the expressions of these measurements and clinicopathologic characteristics. Methods Immunohistochemistry was applied to analyze the expression of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue. The levels of CEA and CA19-9 in serum and clinicopathologic characteristics of all including patients were detected with clinical measurement. All data were analyzed statistically. Results ①The positive rates of Ezrin protein in primary carcinoma of gallbladder and chronic cholecystitis tissue were 66.7% （40/60） and 30.8%（4/13）, respectively (χ2=5.57, Plt;0.05). ②There was no difference between the expression of Ezrin protein in primary carcinoma of gallbladder tissue and age or gender (Pgt;0.05). However, difference was significant between the Ezrin expression and degree of difference, pNevin stages, pTNM stages, lymph node metastasis or distant metastasis (Plt;0.05). ③There were no differences between the positive rates of CEA and CA19-9 in primary carcinoma of gallbladder and age or gender (Pgt;0.05). However, differences were significant between the positive rates of CEA and CA19-9 and pNevin stages, pTNM stages, degree of difference, lymph node metastasis or distant metastasis (Plt;0.05). ④There was some relationship between the expression of Ezrin protein and the positive rate of CEA (rs=0.213, Plt;0.05), but not with the positive rate of CA19-9 (rs=0.081, Pgt;0.05). Conclusions The high expression of Ezrin protein may promote the invasion and metastasis in primary carcinoma of gallbladder. It could be possible to decide the outcome of primary carcinoma of gallbladder through the combined analysis on the expression of Ezrin protein and the serum levels of CEA and CA19-9.
