After the Wenchuan Earthquake on May 12th, 2008, under the b leadership of the SichuanProvincial Party Committee, the People’s Government of Sichuan Province, and the Ministry of Health of the People’sRepublic of China, the Medical Security Team working at the Sichuan Provincial Headquarters for Wenchuan Earthquakeand Disaster Relief Work constructed a secure medical material distribution system through coordination and interactionamong and between regions, systems, and departments.
Objective To investigate the effect of survivin antisense oligodeoxyribonucleotides (survivin ASODNs) on intimal hyperplasia (IH) in vein graft in rats. Methods Autogenous vein graft models were established in 60 Wistar rats by transplanting the interior jugular vein to the common jugular artery using microsurgical technique. The rats were divided into 5 groups according to random digits table, including survivin ASODNs 50 μg group and 200 μg group, scramble ODNs 200 μg group (ODNs group）, Lipofectin+pluronic group and control group. Vein graft samples were collected on 7 d and 14 d after transplantation, respectively. The degrees of hyperplasia were determined and then compared by histomorphology between different groups. The expression of survivin mRNA was measured by RT-PCR and immunohistochemistry. The relevant protein products were detected by Western blot and immunohistochemistry was also used to detect the expression of PCNA. Apoptosis of VSMC was measured by TUNEL.Results Day 7 and 14 were the days that intimal hyperplasied most in control group, ODNs group and Lipofectin+pluronic group, there was no significant difference among these groups yet (Pgt;0.05). The IH could be suppressed by locally transfecting 50 μg of survivin ASODNs (P＜0.05), and it showed a better inhibiting effect in 200 μg of survivin ASODNs group (P＜0.05). The expression of survivin mRNA increased significantly in control group. The expressions of both survivin and PCNA in VSMC significantly decreased in survivin ASODNs group (P＜0.05), whereas the positive cells of TUNEL increased significantly (P＜0.05). Conclusion Transfection of survivin ASODNs may inhibit the IH after vein graft through suppressing the hyperplasia and stimulating the apoptosis of VSMC, and inhibiting the expression of survivin.
To find the relation between the damage of gastric remnant mucosal barrier and the precancerous lesion of gastric remnant mucosa, in the process of the canine gastric remnant precarcinogenesis induced by N-methyN’-nitro-N-nitrosoguanidine (MNNG), we performed regularly the esophagogastroscopy and the mucosal biopsy.At the same time, we also measured gastric transmucosal potential difference and intracellular DNA content of remnant mucosa.We found that the more severe the damage of gastric remnant mucosal barrier was , the greater the malignant capacity of gastric remnant mucosal was.Our study suggests that the damage of gastric remnant mucosal barrier plays an important role in the gastric remnant mucosal precarcinogenesis.
To study the significance of T-lymphocytes rDNA transcription activity in diagnosis, differential diagnosis, therapeutical effect and evaluation of treatment for colorectal carcinoma, 59 cases of colorectal carcinoma, 20 cases of colorectal inflammatory disease and 9 volunteers were choosen to detect the T-lymphocyte rDNA transcription activity of peripheral blood T-lymphocyte by cell culture and CMIAS008 image analysis system of Ag-NOR. Results: T-lymphocytes rDNA transcription activity was decreased obviously in colorectal inflammatory patients. Compared with control group, both group showed markedly statistical difference (Ｐ＜0.01). Tlymphocytes rDNA transcription activity increased gradually to normal groups after operation and chemical treatment for colorectal carcinoma patients; but it decreased for recurrent patients three years after operation. Conclusions: The detection of T-lymphocytes transcription activity can be used as a differential criterion for colorectal carcinoma and colorectal inflammatory disease, meanwhile it also can be used as a reference criterion for evaluation of treatment and supervision of tumor recurrence.
The aim of the this study was to search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture by NP-PCR technique. Bacterial gene fragments were amplified in vitro from DNA which were extracted from cholesterol gallstones in gallbladder for identifying the existence of bacteria. The gallbladder gallstones of 30 patients were analysed. Bacterial DNA was found in the stones of 26 patients, indicating that most cholesterol gallstones harbor bacterial DNA.
The oncogene ras p21 expression and DNA content in 46 cases of colorectal tumor were analysed quantitatively with flow cytometry and cyto-immunofluorescence staining technique. The results showed that the positive rate of ras p21 expression was 65.7% and the rate of DNA aneuploid was 74.3% in colorectal carcinomas. Ras p21 expression was higher in colorectal adenocarcinomas than that of the adenomas and normal mucosa. DNA ploid and proliferative index had some association with ras p21 expresssion. Detection of ras p21 expression and DNA content in tumors may be helpful in predicting the outcome of colorectal cancer patients.
The DNA content, cellular ultrastructure and the expression of blood group Y antigen and immunosuppressive acidic protein-2(IAP-2) were observed in normal breast, cystic hyperplasia of breast and breast cancer. The results showed: the results observed in the cells of cystic hyperplasia with epithelial proliferation grade Ⅰ were similar to those in normal breast cells. The DNA content increased, the hypoplasia and dedifferentiation features in some structures of cellular membrane and nucleus were observed, and the abnormal antigens expressed in part of the atypical hyperplasic cells. The DNA content and ultrastructure in a part of cells with aypical hyperplasia grade Ⅲ were similar to those in the cells of breast cancer grade Ⅰ. The results indicated that in the couse of atypical hyperplasia, the biological abnormalities and its extent of those cells were closely related to the differentiation extent, the developing tendency and the risk of canceration of the cystic hyperplasia of breast.
Dysplasia of gastric stump mucosa in 47 cases was studies.Nuclear DNA contents were measured with an automatic imagie analysis system.The results showed that the mean values of the nuclear DNA contents,area,perimeter,maximum diameter,minimum diameter increased with the increase of severity of dysplasia in gastric stump mucosa(Plt;0.01);where as nuclear form factor decreased with the increase of severity of dysplasia in gastric stump mucosa(Plt;0.05).Severe dysplasia is similar to that of gastric stump cancer in the DNA ploidy histogram.Our results indicate biological behaviour of gastric stump mucosa dysplasia.This study suggests that DNA contents analysis may be used as an important reference for grading,screening,and treating dysplasia of gastric stump mucosa.
Objective To construct small interfering RNA（siRNA） eukaryotic expression vector specific for human hnRNP K gene，and to observe its silencing effects on hnRNP K gene in A549 cells.Methods The expression vectors of pSUPER/hnRNP K siRNAa，pSUPER/hnRNP K siRNAc and pSUPER/siRNAn were constructed by gene recombination and then transfected into the A549 lung carcinoma cell line by using Lipofectamine2000（a and c respectively represented A and C fragments in hnRNP K coding sequence contained 19 nts，n represented nonsense fragment as control）.The mRNA and protein were harvested after 24 h and analyzed for the expression of hnRNP K by RT-PCR and Western blotting respectively.Results The siRNA vector targeted to hnRNP K successfully decreased hnRNP K mRNA and protein levels 24 h after transfection in A549 cells.Relative expressed doses of hnRNP K mRNA in lung cancer cells transfected by hnRNP K siRNAa and hnRNP K siRNAc respectively were 0.24±0.53 and 0.28±0.57 after 24 h，which were significantly lower than that in the control group（both Plt;0.01）.The gray scale values of hnRNP K protein were 0.23±0.11 and 0.28±0.09 respectively，which were also significantly lower than those in the control group（both Plt;0.05）.And pSUPER/hnRNP K siRNAa was the most effective one.Conclusion Eukaryotic expression vector of siRNA specific for hnRNP K is successfully constructed，which lays the basis for the function study of hnRNP K gene and its application in the treatment of lung carcinoma.
Objective To observe the effects of cigarette smoke extract ( CSE) on the proliferation and secretion of hydrogen peroxide ( H2O2 ) in human lung fibroblasts ( HLFs) induced by transforming growth factor-β1 ( TGF-β1 ) . Methods Cultured HLFs were divided into a normal group and a model group induced by TGF-β1 ( 5 ng/mL) , then intervened with CSE at different concentrations ( 0% , 2. 5% , 5% ,10% , respectively) . Brdu ELISA assay was used to detect cell proliferation. H2O2 release from cultured cells was assayed using a fluorimetric method. Cellular localization of H2O2 and expression of α-SMA were performed using a fluorescent-labeling strategy. Results TGF-β1 stimulated group showed positive expression of α-SMA, implying TGF-β1 had induced fibroblasts to differentiate into myofibroblasts. In TGF-β1 stimulated group, 2. 5% and 5% CSE promoted cell proliferation ( P lt; 0. 01 or 0. 05) , while 10% CSE inhibited cell proliferation ( P lt; 0. 01) . In the normal group, both low and high concentration of CSE inhibited cell proliferation ( P lt; 0. 01 or P lt; 0. 05) , and the inhibition effect was dose-dependent. HLF induced by TGF-β1 generated low constitutive levels of extracellular H2O2 that was markedly enhanced by CSE treatment ( P lt; 0. 01) . Pretreatment with DPI, an inhibitor of NADPH oxidase, abolished secretion of H2O2 . Cellular localization of H2O2 by a fluorescent-labeling strategy demonstrated that extracellular secretion of H2O2 is specific to the myofibroblast. Conclusions Low concentration of CSE can promote myofibroblast proliferation, and markedly increase extracellular secretion of H2O2 . CSE possibly take part in the development and progress of idiopathic pulmonary fibrosis by increasing oxidative stress.