【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.
OBJECTIVE: To explore the effects of nandrolone phenylpropionate (NP) on the expression level of pro alpha 1 (I) collagen after burn in rats and the possible mechanism involved in the process. METHODS: Thirty-two Wistar rats with a deep second-degree scald injury and 20% of total body surface area were randomly divided into two groups to receive either 5 mg/kg NP(NP group) or normal saline (control group) every other day. We analyzed the mean integrated optical density(mIOD) of androgen receptor (AR) to determine the distribution and expression of AR in fibroblasts by immunohistochemistry, and measured expression level of pro alpha 1 (I) collagen mRNA by quantitative fluorescent RT-PCR to find the relation between expressions of AR and pro alpha 1 (I) collagen mRNA. The total specimens were obtained from the scalded rats after 4, 7, 14 and 21 of after burn. RESULTS: The expression of pro alpha 1 (I) collagen mRNA in NP group was significantly higher than that in control group on the 7th, 14th and 21st days(P lt; 0.05), but there was no significant difference on the 4th day. The density of AR in fibroblasts had significant difference (P lt; 0.05) between the two groups after 4, 7, 14 and 21 days. A positive relationship existed between the expression of pro alpha 1 (I) collagen mRNA and quantity of AR in fibroblasts(r = 0.836). CONCLUSION: The nandrolone phenylpropionate increased the expression of pro alpha 1 (I) collagen mRNA and enhanced the density of AR in fibroblasts. The higher expression of pro alpha 1 (I) collagen mRNA had a relation with the change of quantity of AR in fibroblasts.
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.
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.
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.
Objective To study the significances and expressions of Y14 and Upf1 in human breast cancer tissues and human normal breast tissues. Methods The expressions of Y14 and Upf1 were determined in human breast cancer tissues and human normal breast tissues by immunohistochemistry and laser scanning confocal microscope. Results ①Y14 and Upf1 levels of human breast cancer tissues were obviously higher than those of human normal breast tissues (Plt;0.05). ②Y14 and Upf1 levels of the class Ⅲ were obviously higher than those of the class Ⅰ and Ⅱ in breast cancer tissues (Plt;0.05). ③Y14 and Upf1 levels in breast cancer tissues of patients with positive axillary lymph nodes were obviously higher than those with negative axillary lymph nodes (Plt;0.05). Conclusion The function of nonsense-mediated mRNA decay (NMD) is obviously enhanced in breast cancer tissues. Expressions of Y14 and Upf1 levels of the class Ⅲ cancer tissue and cancer with positive axillary lymph node are high because of NMD.
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 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 study the protective effects of pre-storing glycogen on warm ischemia reperfusion injury during partial hepatectomy. Methods Thirty-eight patients were randomly divided into a trial group (n=19) and a control group (n=19). In the trial group, patients were given high concentration glucose intravenously during the 24 hours before the operation. The hepatic lesion was resected after portal triad clamping in the two groups. Liver function of all patients was measured before the operation and the first and fifth days after the operation. Normal hepatic tissue was biopsied to measured hepatic tissue glycogen contents before the operation and the change of superoxide dismutase (SOD) at the point of pre-ischemia, post-ischemia, and reperfusion 2 hour. Bcl-2 mRNA, a well known anti-apoptotic factor, was also detected using quantitative polymerase chain reaction. Results The hepatic tissue glycogen content of the trial group was significantly higher than that of the control group before the operation (Plt;0.01). Liver function of the trial group was significantly better than that of the control group on the first and fifth day after operation (Plt;0.05). There was significant difference in SOD activity between the two groups at the end of hepatic vascular occlusion and at the point of 2-hour reperfusion (Plt;0.05). Furthermore Bcl-2 mRNA expression of the trial group was notably up-regulated at the point of 2-hour reperfusion compared to the control group. Conclusion Pre-store storing glycogen might protect liver ischemia reperfusion injury caused by hepatic vascular occlusion during partial hepatectomy. The potential mechanism might be that pre-storing glycogen enhances Bcl-2 expression.
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.
