【Abstract】ObjectiveTo study the expressions of matrix metalloproteinase 2 (MMP2) and carbohydrate antigen 50 (CA50) in colorectal carcinoma, cancer-adjacent mucosa （2 cm from the nether edge of tumor）, cancerdistant mucosa （5 cm from the nether edge of tumor） and normal colorectal mucosa, and to elucidate their effects on the development of colorectal carcinoma. MethodsThe expressions of MMP2 and CA50 were detected immunohistochemically in 40 cases of colorectal carcinoma, cancer-adjacent mucosa, cancer-distant mucosa and 10 cases of normal colorectal mucosa. Results①The expression intensity and positive rates of MMP-2 and CA50 increased significantly in turn by normal mucosa, cancer-distant mucosa, cancer-adjacent mucosa and colorectal carcinoma. ②The expression of MMP2 was correlated with CA50 in colorectal carcinoma. ③The expression of CA50 in colorectal carcinoma was closely associated with tumor differentiation, and the expression of MMP2 in colorectal carcinoma was closely associated with differentiation and Dukes stages as well. ConclusionOver expression of MMP2 facilitates the malignant progress of colorectal carcinoma; CA50 is a reliable marker of malignance in colorectal carcinoma; CA50 and MMP2 may have synergetic effects on the development of colorectal carcinoma.
Objective To observe the inhibitory effects of local co-transfection of tissuetype plasminogen activator(tPA) gene and proliferating cell nuclear antigen antisense oligodeoxynucleotides(PCNA-ASODN) on the intima proliferation and restenosis of autograft artery in rabbits. Methods One hundred and twenty male Zelanian rabbits were randomly divided into four groups(n=30, in each group): control group, PCNA-ASODN group, tPA group and tPA+PCNAASODN group. The left and right external iliac arteries (length 1.0 cm) were transplanted reciprocally. The transplanted arteries were respectively soaked in lipofection, PCNAASODN, pBudCE4.1/tPA and pBudCE4.1/tPA+PCNA-ASODN solution about 15 minutes. The transplanted arteries were sutured with 9-0 sutures soaked in PCNA-ASODN and pBudCE4.1/tPA solution. Each group were divided into five subgroups(n=6, in each subgroup) according to the sacrifice time (3 d, 7 d, 14 d, 28 d and 56 d after operation). On every sacrifice time point, the vascular specimens were harvested. The thrombocyte assembling and thrombus forming lining vessel wall were observed by scanning electron microscope. The pathological morphology of transplanted arteries were observed under microscope(HE). The intimal areas and stenosis ratio(%) of transplanted arteries were calculate and analyzed statistically among groups by computer system. The mRNA expression of tPA gene in transplanted ressel wall was detected with vevere transcriptionPCR(RT-PCR). The number of PCNA positive cells in transplanted vessel wall was counted by SP immunochemisty.Results The mRNA expression of tPA gene in the transplanted vessel wall in tPA and tPA+PCNA-ASODN groups was higher than that of the other two groups(P＜0.01).The number of PCNA positive cells in the transplanted arteries in PCNAASODN, tPA and tPA+PCNAASODN groups were significantly lower than that of control group(P＜0.05，P＜0.01). The intimal areas and degrees of luminal stenosis of PCNAASODN, tPA and tPA+PCNAASODN groups were lower than those of control group(P＜0.05，P＜0.01), and those of tPA＋ PCNA-ASODN group were lower than those of PCNA-ASODN and tPA groups(P＜0.05). Scanning electron microscopy showed that there were a few thrombocytes lining the vessel wall of tPA group and tPA+PCNAASODN group and no thrombus, whereas there were abundant thrombocytes and thrombi lining the vessel wall of the control group. Conclusion Co-transfection of tPA gene and PCNA-ASODN can effectively inhibit the proliferation of VSMC, hyperplasia of intima and restenosis of transplanted artery.
Objective To investigate the effects of recombinant adenovirus-mediated co-transfection of carcinoembryonic antigen (CEA) gene and erythropoietin (EPO) gene on promoting hematopoietic stem cells directly producing erythrocyte vaccine against colon cancer. Methods The expression adenovirus vectors carrying CEA and EPO or green fluorescent protein (GFP) gene were constructed respectively, and recombinant adenovirus carrying CEA, EPO or GFP were packaged and produced respectively. The bone marrow-derived mesenchymal stem cells (MSCs) of mice were isolated and cultured in vitro by anti-CD117 magnetic bead separation, and were transfected with CEA (CEA group), EPO (EPO group) or GFP (blank vector group), co-transfected with CEA and EPO (CEA-EPO group). The expressionsof CEA and EPO gene and its protein after transfection in supernatant fluid of culture were detected by realtime-PCR and Western blot method in each group. We had checked and obtained the vaccine with co-transfection of CEA gene and EPO gene by cell red line marker antibody CD71 and GPA, then we carried on experiments with the vaccine in vitro and in vivo. There were 4 groups in our trail: blank vector group, CEA group, EPO group, and CEA-EPO group. Results We had successfully gathered the hematopoietic stem cells, flow cytometry analysis result showed that there were significant differences before and after purification for positive selected samples (P<0.05). The expressions of double genes (CEA-EPO gene) and protein showed CEA-EPO gene were successfully transfected into the hematopoietic stem cells. We had confirmed erythrocyte vaccine with co-transfection of CEA and EPO gene by antibody CD71 and GPA with flow cytometry. The monocytes cytotoxicity on colon cancer cell line CT26 showed that lysis of target cells of CEA-EPO group were higher than those of other 3 groups when in proportion of 40∶1 (P<0.05). In the experimentation of neoplasma format, the volume of tumor and mortality were smaller or lower, but survival time was longer of CEA-EPO group in2 weeks after treatment (P<0.05). Conclusions The erythrocyte vaccine with co-transfection of CEA gene and EPO gene has efficient anti-tumor effects on colon cancer. Not only can promote hematopoietic stem cell directly producing erythrocyte vaccine, but also can produce tumor antigen vaccine against colon cancer.
The expression of T antigen in rectal cancer and mucosa remote from carcinoma by immunohistochemistry was investigated. Mucin protein was also examined by HID-AB staining. The results showed that the expression of T antigen in rectal cancer was much ber than those in 10cm mucosa remote from carcinoma and no significant difference as compared with 5cm mucosa. The sialomucin reactions in 5cm and 10cm mucosa remote from carcinoma were 45% and 20% respectively. The coincident sialomucin positive reaction and expression of T antigen were found in 40% 5cm remote mucosa .There is significant correlation between them (Ｐ＜0.05). The authors conclude that the expression of tumorrelated antigen and change of mucin protein in remote mucosa without malignant invasion may suggest the malignant potential of the mucosa. Further investigations should be performed into the effect of these changes on the local recurrence after redical resection of rectal cancer.
【摘要】 目的 检测B细胞成熟抗原（BCMA）mRNA在系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）的表达水平，探讨BCMA在SLE发病中的意义。 方法 纳入2006年1-11月收治的36例SLE患者，同期17例健康志愿者作为对照组，采用半定量RT-PCR法检测外周血单个核细胞中BCMA mRNA的表达，并与SLE疾病活动指数（SLEDAI）进行相关性分析。 结果 SLE患者组BCMA mRNA表达水平（0.598±0.230）均明显高于正常对照组（0.411±0.309）（Plt;0.05）。SLE患者BCMA mRNA表达水平与SLEDAI评分无相关性（P=0.590）。 结论 SLE患者BCMA mRNA表达水平的增高，可能在SLE的发病机制中具有一定的作用。【Abstract】 Objective To detect the mRNA expression of B-cell maturation antigen (BCMA) in peripheral blood mononuclear cells (PBMC) in patients with systemic lupus erythematosus (SLE), and explore the role of BCMA in the pathogenesis of SLE. Methods From January 2006 to November 2006 the expression of BCMA mRNA in PBMC of 36 patients with SLE and 17 normal controls were measured by half-quantitative RT-PCR. The linear correlation between the expression of BCMA mRNA and SLE disease activity index (SLEDAI) was assessed. Results The level of BCMA mRNA (0.598±0.230) in PBMC significantly increased in SLE patients compared with that in the normal controls (0.411±0.309) (Plt;0.05). The expression of BCMA mRNA in SLE patients showed no correlation with SLEDAI score (P=0.590). Conclusion The results suggest that the expression of BCMA mRNA might play an important role in the pathogenesis of SLE.
Insufficient supply of organ for allotransplantation made the study on finding new organ resources from animal progress. Pig is regarded as one of the optimal donor animals for human. The major obstacle in this field is hyperacute reaction (HAR), which is triggered after the xenogenic natural antibodies preexisting in recipient blood combine to the antigens on the surface of the endothelium and activate the complement system. alpha-Galactose residues (alpha-Gal) on the endothelial cell have been identified as the major xenoantigens. NJZ Pig has been closely breed since 1938, whose family history is clear. Tissue samples from heart, liver, kidney, pancreas, lung, small intestine, skin, spleen, thymus and lymph node were obtained and embedded in paraffin. The sections were performed the immunohistochemical staining with the sera from health volunteers (including all the blood types) as the primary antibodies as well as the biotin labeled bandeirae simplicifolia I isolectin B4 (BS I-B4), which has specific affinity to alpha-galactose. All the staining sections were compared with the tissues digested with alpha-galactosidase. There was no difference between the antigens recognized by sera of different blood types. alpha-Gal was still the major xenoantigen on the endothelial cells. There might exist non-alpha-Gal antigens on the distal convoluted tubules and collecting tubules of the kidney. There was no alpha-Gal distributing on the secreting part of pancreas, either the islet cells or the matrix cells, but surely on pancreatic duct and vessels. All the antigenity was destroyed after the enzyme digestion except that the small intestine gland still positive with the BS I-B4. alpha-Gal is the major xenogenic antigen in NJZ Pigs. There exist some unknown antigens on the distal convoluted tubules and collecting ducts of the kidney. The blood type of recipient is not the first affair to be considered in pig-to-human xenotransplantation. The specificity of BS I-B4 for the alpha-galactose needs more detail research.
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)
ObjectiveTo explore value of preoperative neutrophil-to-lymphocyte ratio (NLR) combined with carbohydrate antigen 19-9 (CA19-9) in evaluating prognosis for patients with pancreatic head carcinoma.MethodsThe clinical data of patients with pancreatic head carcinoma underwent radical pancreaticoduodenectomy in the Lanzhou University Second Hospital from February 2011 to August 2017 were retrospectively analyzed. The preoperative serum albumin (ALB), CA19-9, NLR, carcino-embryonic antigen (CEA), alpha-fetoprotein (AFP), N stage, tumor differentiated degree, and other indicators were recorded and the patient’s overall survival was recorded. Linear correlation analysis was used to examine the association of NLR with serum CA19-9. The receiver operating characteristic (ROC) curve was used to calculate the cutoff values of NLR and CA19-9 corresponding to the overall survival rate of 2-year. Kaplan-Meier method was used for survival analysis. Cox proportional hazards regression model was used to evaluate the prognostic value of preoperative CA19-9 and NLR.Results① A total of 121 patients were enrolled in this study. There was no significant correlation between the preoperative serum CA19-9 and NLR (r=0.100 05, P=0.272 9). ② Multivariate analysis showed that higher NLR, higher CA19-9 level, higher N stage, and lower tumor differentiation were the risk factors for overall survival of pancreatic head carcinoma (P<0.05). ③ The cutoff values of CA19-9 and NLR in evaluating the 2-year overall survival rate were 123.3 U/mL and 2.34 respectively by the ROC curve analysis, the area under ROC curve of CA19-9, NLR, and CA19-9 combined with NLR were 0.763, 0.686, and 0.823, respectively. ④ The proportions of patients with CEA≥5 μg/L and ≥N1 stage were higher in the patients with CA19-9≥123.3 U/mL as compared with patients with CA19-9<123.3 U/mL (P<0.05); The proportions of patients with preoperative serum ALB level <35 g/L and CEA≥5 μg/L, ≥N1 stage, and poor differentiation were higher in the patients with NLR≥2.34 as compared with the patients with NLR<2.34 (P<0.05). ⑤ Kaplan-Meier survival analysis showed that patients with CA19-9≥123.3 U/mL and NLR≥2.34 had worse overall survival than those patients with CA19-9<123.3 U/mL and NLR<2.34 (HR=3.599, P<0.01; HR=2.506, P<0.01). The patients with CA19-9≥123.3 U/mL and NLR≥2.34 (n=42), CA19-9≥123.3 U/mL or NLR≥2.34 (n=48), CA19-9<123.3 U/mL and NLR<2.34 (n=31) had better overall survival in turn (overall and between groups, all P<0.01).ConclusionPreoperative serum CA19-9 combined with NLR has an important value in evaluating prognosis for patients with pancreatic head carcinoma.