【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
ObjectiveTo explore feasibility of mRNA vaccines as a novel strategy for individualized precision treatment of pancreatic cancer (PC). MethodThe recent domestic and international literature on vaccine research in PC was reviewed. ResultsThe heterogeneity between and within the pancreatic tumors had limited the efficacy of traditional vaccines based on cells, exosomes, proteins, peptides, or DNA for PC. The mRNA vaccine was considered as a promising alternative therapy due to its precise targeting, low toxicity, and ability to induce long-lasting immune memory. Breakthroughs in the tumor antigen recognition, immune subtype differentiation, and mRNA vaccine construction, the development strategy of PC mRNA vaccine would further facilitate the development of personalized precision medicine. The existing mRNA vaccines usually need to be combined with other immunotherapy methods to improve efficacy, while the development of preventive vaccines is still exploraing. ConclusionsmRNA vaccines, as an innovative and promising platform, offer a new hope for the development of PC vaccines. However, the heterogeneity of PC has resulted in poor efficacy with traditional vaccines. Although the limitations of traditional vaccines and the heterogeneity of PC itself, the more challenges of vaccine research of PC is facing, the advantages of mRNA vaccine still make it possible to treat PC. In the face of the challenge of complex characteristics of PC, more research is needed to support the transformation and application of mRNA vaccine in clinical therapy.
【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.
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.
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 study the mechanism of nandrolone phenylpropionate (NP) on hepatic albumin mRNA and androgen receptor(AR) in burned rats. Methods Thirty-two Wistar rats with a deep second-degree cutaneous burn of 20% total body surface area were randomly divided into two groups:NP group (experimental group, 5 mg/kg NP) and normal saline as control group every other day. The expression copy quantity of albumin-mRNA and mean integrated absorbency(mIA) of AR in liver tissue were measured by quantitative fluorescent RT-PCR and immunohistochemistry respectively on the 4th, 7 th, 14th and 21st days of post-burned. Result The expression levels of ablumin-mRNA and AR in liver tissue in HP grouop were much higher than those in control group. The ablumin-mRNA and AR expression increased significantly(P lt; 0.05) after 7 and 14 days, whole the expression had no significant difference between NP group and control group on the 4th day. A positive correlation occurred between the expression level of albumin-mRNA and the quantity of AR in liver tissue(r=0.936, P lt; 0.05). Conclusion Nadrolone phenylpropionate up-regulated respectively the expression of albumin-mRNA and the density of AR in liver tissue.
Objective To investigate the percentage of CD4 + CD25 + Treg cells and expression of Foxp3 mRNA in asthmatic patients and the impacts of inhaled steroids.Methods The percentages of CD4 +CD25 + Treg cells was assayed by flow cytometry and the expression of Foxp3 mRNA was detected by RT-PCR in peripheral blood mononuclear cells from the patients with chronic persistent asthma before and after steroids inhalation in comparison with healthy control. The forced expired volumin one second/predicted value( FEV1% pred) and peak expired flow( PEF) were measured by spirometry. Results The level of CD4 + CD25 + Treg cells and the expression of Foxp3 mRNA were lower in asthmatics before steroids treatment than those in control ( P lt; 0. 05) which were increased significantly after steroids treatment ( P lt; 0. 05) .FEV1% pred and PEF were declined significantly than those in control but improved markedly after treatment ( P lt; 0. 05) . Conclusions The insufficiency of amount and function of immue-suppressive CD4 + CD25 +Treg cells may play a role in the pathogenesis of asthma. Inhaled steroids can improve the lung function of asthmatics by upregulating the level of CD4 + CD25 + Treg and Foxp3.
Objective To investigate the relationship between gene expression of endothelin-3 (ET-3） and inflammation of acute pancreatitis （AP） in rats. Methods Fifty-four rats were divided randomly into 4 groups: sham operation group, AP group, arterial injection group and vein injection group. AP was induced by reverse intra-bile duct infusion 4.5% sodium taurocholate, treated with low dose dopamine 〔5 μg/（kg·min）〕 by injecting arterial or tail vein. Rats were sacrificed at 1， 6 and 24 h after the induction of AP. The mRNA expression of ET-3 was evaluated by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and pathological changes was observed in rats. Results Expression of ET-3 mRNA could be detected from 1 up to 24 h after the induction of pancreatitis. Expression of ET-3 mRNA of sham operation group was decreased significantly compared with other three groups. Expression of ET-3 mRNA showed a significant decrease by arterial injection dopamine than that by tail vein (P＜0.05， P＜0.01). The pathologic score in AP group was the highest, vein injection group was the next one, and score in sham operation group was the lowest. Conclusion There are significant relationship between inflammation of AP and expression of ET-3 mRNA. Dopamine administration by arterial injection is more effective than that by tail vein injection.
【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.