ObjectiveTo test the expressions of human mammary gland globin (hMAM) mRNA in the peripheral blood of breast cancer and breast benign lesions patients, try to provide the theory basis for the choice of breast cancer molecular marker. MethodsPolymerase chain reaction (PCR) technology was used to detect the expressions of hMAM mRNA in peripheral blood of 78 cases of breast cancer patients, 15 cases of hyperplasia of mammary gland, and 15 cases of breast fibroadenoma. The relationship between the expressions of hMAM-mRNA in peripheral blood of breast cancer patients with patient's age, tumor size, pathological type, tumor stage, axillary lymph node metastasis, and the ER, PR and HER-2 status were analyzed. ResultsThe expressions of hMAM-mRNA in peripheral blood were not detected in breast hyperplasia and breast fibroadenoma patients, but the peripheral blood hMAM-mRNA expression rate in breast cancer patients was 48.72% (38/78), the difference was statistically significant (χ2=12.357, P=0.000). The expression of peripheral blood hMAM mRNA was not related to the patient's age, tumor size, pathological type, and ER, PR and HER-2 status (P > 0.05), but the expression of peripheral blood hMAM mRNA was related to the clinical staging of tumor (Z=-2.214, P=0.027) and lymph node metastasis status (Z=-2.754, P=0.006). ConclusionPeripheral blood hMAM-mRNA detected is a sign of breast cancer, further research is needed to confirm whether hMAM mRNA detection in peripheral blood correlates with poor prognosis of breast cancer patients.
Objective To investigate the effects of expression of TNFα mRNA on glucose uptake in both the liver and skeletal muscle after endotoxemia. Methods In the mice with intraperitoneal injection of lipopolysaccharide (LPS), the changes of TNFα level of plasma and uptake of 2-deoxyglucose (2-DG) in the isolated soleus muscle and hepatic tissues were determined, then the reinstatement of glucose uptake by injecting TNF-McAb for 3 days was also observed. In addition, changes of TNFα mRNA expression of liver were evaluated. Results The expression of TNFα mRNA in the liver showed markedly increased in the first 3 hours post endotoxemia and remaind high for 3 days, and the plasma TNFα level paralleled with TNFα mRNA expression of liver also was elevated. The basal uptake of 2-DG both in muscle and liver were markedly increased, but the stimulated 2-DG uptake with insulin was greatly reduced as compared with the control. In addition, these abnormalities of 2-DG uptake can be partially corrected by neutralization of the circulatory TNFα by administration of TNF-McAb. Conclusion The disorders of glucose uptake of the liver and the muscle due to the overexpression of TNFα mRNA and elevated circulatory TNFα level may be the mechanism of insulin resistance after endotoxemia.
Abstract： Objective To evaluate the sensitivity, specificity and clinical significance of Lunx mRNA in surveying micrometastasis by sampling peripheral blood of lung cancer patients, studying the early diagnosis of lung cancer metastasis. Methods From March 2004 to February 2005，Reverse transcriptionpolymerase chain reaction(RT-PCR) was used to detect Lunx mRNA of peripheral blood of 60 lung cancer patients（lung ancer group）. Peripheral blood of 20 patients with pulmonary benign lesions (pulmonary benign lesions group) and 10 normal healthy volunteers (control group) were used as control. Results (1) In the lung cancer group, Lunx mRNA were expressed positive in 28(46.7％) patients. All the pulmonary benign lesions group (0/20) and the control group (0/10) were expressed negative. (2) One of the 12 stage I patients with lung cancer (8.3％) was positive for Lunx mRNA, 5 of the 15 stage Ⅱ patients (33.3%) were positive, 22 of the 33 stage Ⅲ patients (66.7%) were positive. Comparing the positive rate of these groups, there was no statistically difference between stage Ⅰ and stage Ⅱ, but the difference between stage Ⅰ+ stage Ⅱ and stage Ⅲ significant (χ2=15.88, P=0.000). (3) In 38 adenocarcinoma, 17 were positive for Lunx mRNA. In 14 squamous carcinoma, 7 were positive. All the 3 adenosquamous carcinoma expressed positive. 1 of 3 small cell lung cancer was positive, 1 large cell carcinoma and 1 carcinoma sarcomatodes expressed negative. Comparing the positive rate of these groups, there was no statistically difference among them. (4) By followup till March 2005, 10 lung cancer patients were found metastasis. Among them, 9 were positive for Lunx mRNA expression, and 1 was negative. Conclusion Lunx mRNA has high sensitivity and specificity in surveying micrometastasis by ampling peripheral blood. It would likely to be an proper gene for the detection of micrometastasis in lung cancer patients.
【Abstract】Objective To explore the changes of expression of AFP mRNA in human hepatocellular carcinoma （HCC） tissues after oral Xeloda therapy.Methods Total RNA was extracted from HCC tissue samples collect after operation and nested reverse transcription polymerase chain reaction （RT-nested PCR） assay was performed to determine the expression of AFP mRNA in this study.Results The final product of AFP mRNA amplified by RT-PCR was 174 bp and by RT-nested PCR was 101 bp. The AFP mRNA is positive in 12 of 21 patients (positive rate 57.14%) amplified by RT-nested PCR assay in Xeloda treatment group which is much lower than control group: 18 of 20 patients (positive rate 90.00%),P＜0.05.The serum AFP value of Xeloda treatment group 〔(23.2±12.8) μg/L〕 is much lower than that of control group 〔(39.6±24.3) μg/L〕 four weeks after operation (P＜0.05). However, There was no difference between two groups in serum AFP value before operation.Conclusion Xeloda can effectively suppress the expression of AFP mRNA in human HCC tissues and lower it’s product serum AFP value.The clinical application of Xeloda in HCC patients deserve further study.
OBJECTIVE: To elongate the proliferation life-span of human umbilicus vein endothelial cell (HUVEC). METHODS: We synthesized the human telomerase reverse transcriptase mRNA (hTERT mRNA) by in vitro transcription, then transferred the hTERT mRNA into HUVEC in quicent stage by lipofect introduction. RESULTS: Telomerase expressed transiently in HUVEC, and the cell life-span was elongated for 7 population doublings. CONCLUSION: Telomerase can be reconstructed controllably and transiently in HUVEC by hTERT mRNA introduction, this method has the potential to be used to elongate the lifespan of cells cultured in vitro.
【Abstract】Objective To investigate whether liver resection for hepatocellular carcinoma (HCC) causes dissemination of liver tumor cells into blood circulation. Methods Fourteen patients with HCC, but without evidences of metastasis, were enrolled for the study. Blood samples of peripheral blood before skin incision and after abdominal wall suture, and of hepatic venous blood and portal venous blood after liver parenchyma dissection, were obtained. AFPmRNA was detected by reverse transcription polymerase chain reaction assays, the change of the level of its expression during operation was assessed by semi-quantitative analysis. Results The rate of its expression before and after operation in peripheral blood, and during operation in portal venous blood and in hepatic venous was 42.9%, 35.7%, 42.9% and 57.1% respectively. There were no differences between them. However, the level of its expression in hepatic venous blood was significantly higher than others (P＜0.05). Conclusion Liver resection for HCC induces releases of cells from the liver, probably including tumor cells, into blood circulation.
【Abstract】ObjectiveTo investigate the expression of Tob mRNA in human colorectal cancer tissues, and their corresponding paracancerous normal tissues which was 10 cm above the tumor and pathologically proved and to explore the role of Tob mRNA in the pathogenesis of colorectal cancer. MethodsQuantitative real time RTPCR was used to detect the expression of Tob mRNA in 31 colorectal cancers. ResultsCompared with paracancerous tissue, the expression of Tob mRNA in colorectal cancer tissues was significantly increased. Moreover, the expression levels of Tob in Dukes A, B, C, D were 1.146±0.067, 1.120±0.073, 1.052±0.020 and 1.047±0.010 respectively. Analyzed by oneway ANOVA, there were significant differences in expression of Tob in different Dukes stage. ConclusionThe upregulation expression of Tob mRNA may be closely associated with tumorigenesis of colorectal carcinoma.
Objective To detect expression of CK20 mRNA in peripheral blood of patients with colorectal carcinoma and its clinical significance. Methods Using the reverse transcriptase-polymerase chain reaction (RT-PCR),CK20 mRNA expression was examined in peripheral blood from 42 patients with colorectal carcinoma before and after operation, 20 healthy volunteers, 20 fresh colorectal carcinoma samples. Results The positive expression rates of CK20 mRNA were 45.24%(19/42) and 33.33%(14/42) before and after operation in 42 colorectal carcinoma patients respectively. All 20 fresh colorectal carcinoma samples revealed expression of CK20 mRNA, but the 20 normal blood samples did not. Conclusion The detection of CK20 mRNA in peripheral blood is helpful to early diagnose, assess the prognosis and make a correct treatment of colorectal carcinoma.
Objective To observe the expression of proinflammatory factors messenger RNA(mRNA) in periretinal membrane of proliferative vitreoretinopathy. Methods Fourteen specimens of periretinal membrane were collected during vitrectomy, and they were made into paraffin sections.The presence of mRNA coding for IL-1,IL-6,IL-8 and TNF alpha was observed by in situ hybridization(ISH) with biotin labeled oligonuclotide probes respectively.The eyeball after corneal grafting was made as normal control. Results No expression of proinflammatory factors mRNA was found in normal human retina.Positive staining was present in 5 specimens.In these specimens, IL-1βmRNA was found in 3 specimens and TNFαmRNA was found in 3 specimens,there is 1 specimen expressed IL-8 mRNA and 3 specimens expressed IL-6 mRNA.In these positive specimens, one contained cells expressing mRNA for IL-1βbeta and IL-6, and one exhibited cells expressing mRNA for IL-1β、IL-8 and TNFα,two membranes possessed positive cells for IL-6 and TNFαmRNA, one membrane contained IL-1βmRNA positive cells only. Conclusion These findings suggest that these cytokines may be locally produced by cells infiltrating epiretinal membranes. The expression of IL-1β, IL-6, IL-8 and TNFαmRNA within retinal membranes provides further evidence of a pathogenic role of these cytokines in proliferative vitreoretinopathy. (Chin J Ocul Fundus Dis, 2002, 18: 286-288)
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.