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1.
Journal of Chongqing Medical University ; (12)2007.
Article in Chinese | WPRIM | ID: wpr-580144

ABSTRACT

Objective:To investigate the differential expression of VEGF165b in the transitional cell carcinoma of the bladder (TCCB) and normal bladder tissues and its role in the development of TCCB. Methods: S-P immunohistochemistry was used to detect the expression of VEGF165b protein in 38 specimens of paraffin-embedded TCCB tissues and 36 specimens of paraffin-embedded normal bladder tissues. RT-PCR was performed to detect the expression of VEGF165b mRNA in 55 specimens of fresh TCCB tissues and 43 specimens of fresh normal bladder tissues. Results: Among 36 of normal bladder tissues, 35 specimens had positive expression of VEGF165b protein, with the positive rate of 97.22% (35/36) that was significantly higher than 21.05% (8/38) (P

2.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 298-300, 2006.
Article in Chinese | WPRIM | ID: wpr-974109

ABSTRACT

@#ObjectiveTo investigate the rule of changes of the soleus mass and expression of myosin heavy chain (MHC) isoforms mRNA.Methods40 female Wistar rats were divided randomly into the control group and three spinal cord transection (ST) groups, ST7, ST15, and ST30 with 10 animals in each group. Rats in ST groups were subjected to a complete ST between T8 and T10 levels. The right soleus was dissected and weighed at 7, 15, 30 days after ST, and the expression of MHC mRNA isoforms was measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).ResultsThe Absolute and relative soleus masses in three ST groups were lower significantly than those of control group (P<0.05). The soleus mass in ST15 and ST30 groups were lower than that of ST7 group (P<0.05). The soleus of control group predominantly expressed MHC-I and some MHC-IIa, whereas the soleus began to express MHC-IIx and MHC-IIb after ST, except for MHC-I and MHC-IIa. ST induced consistently down-regulation of MHC-I mRNA and up-regulation of MHC-IIx and MHC-IIa at three time points after ST. The level of MHC-IIb mRNA expression was very low at three time points after ST.ConclusionST can influence the soleus mass at early stage after ST. ST induces a shift toward a faster muscle phenotype from slow to fast MHC isoform. MHC demonstrates plasticity in response to decrease neuromuscular activation.

3.
Cancer Research and Clinic ; (6)2006.
Article in Chinese | WPRIM | ID: wpr-676677

ABSTRACT

Objective To detect EIA factor expression in human rectal cancer and normal tissue and to determine whether it is correlated with invasion and metastasis of human rectal cancer. Methods Real- time RT-PCR was used to detect E1AF expression in matched rectal cancers and normal tissues from g6 in- patients.Results Among the 86 rectal cancer samples tested,55 cases E1AF mRNA overexpression was ob- served. The mRNA expression of E1AF in the sample group was remarkably different from that in the control group.In carcinomas,E1AF mRNA expression correlated significantly with histological type,depth of inva- sion,lymph node and distant metastasis and advanced Duke stage.Conclusion E1AF is correlated signifi- cantly with invasion and metastasis of human rectal cancer and may be an important factor in the invasion and metastasis.

4.
Korean Journal of Otolaryngology - Head and Neck Surgery ; : 204-207, 2005.
Article in Korean | WPRIM | ID: wpr-649154

ABSTRACT

BACKGROUND AND OBJECTIVES: Salivary secretions and the secreted IgA in the secretions play a critical role in maintaining oral health via innate host defense mechanism. Cathelicidins are a family of peptides thought to provide an innate defensive barrier against a variety of potential microbial pathogens. LL-37, an antimicrobial peptide, is the only Cathelicidin protein so far identified in humans. The purpose of this study was to examine the expression of Cathelicidin in human salivary glands and to investigate upregulation of Cathelicidin in inflammatory conditions. MATERIALS AND METHOD: Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining were performed on 20 salivary gland tissues, of which 10 were normal and 10 were chronic sialadenitis. RESULTS: Cathelicidin mRNA transcripts were detected in the normal salivary glands and chronic sialadenitis. The level of Cathelicidin mRNA in chronic sialadenitis was significantly increased compared with that in the normal salivary gland. Cathelicidin protein was expressed in the glandular epithelium of the normal salivary gland and chronic sialadenitis. CONCLUSION: The results indicate that Cathelicidin might play an important role in the innate host defense of human salivary glands.


Subject(s)
Humans , Cathelicidins , Epithelium , Immunoglobulin A , Oral Health , Peptides , RNA, Messenger , Salivary Glands , Sialadenitis , Up-Regulation
5.
Journal of the Korean Surgical Society ; : 194-198, 2004.
Article in Korean | WPRIM | ID: wpr-172432

ABSTRACT

PURPOSE: Micrometastasis is known as a significant predictor of prognosis in colorectal cancer patients. Recently, reverse transcriptase polymerase chain reaction (RT-PCR) has been applied to detecting micrometastasis. The drainage vein and peritoneum were examined and the micrometastases assessed in a series of colorectal cancer patients. METHODS: 22 patients, who were histologically diagnosed with colorectal cancer, and 8 patients of serosal and peritoneal brushing, were examined using RT-PCR to amplify the mRNAs for two epithelial markers, carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20). RESULTS: Among the 22 colorectal cancer patients, the positive rates of CK-20 and CEA mRNAs in the drainage vein were 10 (45%) and 7 (32%), and those of the serosal and peritoneal brushing were 6 (75%) and 5 (63%), respectively. CONCLUSION: These results suggest that the "no touch isolation technique" might be useful for operations in advanced colorectal cancer patients, and the brushing of the serosal or Douglas pouch can represent the micrometastasis status.


Subject(s)
Humans , Carcinoembryonic Antigen , Colorectal Neoplasms , Douglas' Pouch , Drainage , Keratin-20 , Keratins , Neoplasm Micrometastasis , Peritoneal Cavity , Peritoneum , Prognosis , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger , Veins
6.
Journal of the Korean Gastric Cancer Association ; : 128-133, 2003.
Article in Korean | WPRIM | ID: wpr-132070

ABSTRACT

PURPOSE: The benefits of the "no-touch" isolation technique that is usually performed to prevent the circulation of tumor cells are not evident. The aim of this study was to determine whether the no-touch isolation technique for treating gastrointestinal cancers could prevent the circulation of tumor cells detected by reverse transcriptase polymerase chain reaction (RT-PCR). Matrials and Methods: By using RT-PCR to amplify mRNAs for two specific epithelial markers, carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20), we examined 34 gastric cancer patients who had been histologically diagnosed and 22 patients had undergone serosal and peritoneal brushing. RESULTS: In 10 (29.4%) of the 34 gastric cancer patients, we detected CK20 mRNA before manipulation, and in 17 (51.5%) of those patients, after we detected it. The density of the CK20 mRNA band was increased in 11 cases (33.3%) and the density was decreased in 2 cases (6.1%). In 16 (48.5%) of the 34 gastric cancer patients, we detected CEA mRNA before manipulation, and in 17 (51.5%) patients after we detected it. The density of the CEA mRNA band was increased in 8 cases (24.2%) and decreased in 3 cases (9.1%). CONCLUSION: These result suggest that the "no-touch isolation technique" might be useful when operating on advanced gastric cancer patients and that serosal or Douglas pouch brushing can be used to determine the status of micrometastasis.


Subject(s)
Humans , Carcinoembryonic Antigen , Douglas' Pouch , Gastrointestinal Neoplasms , Keratin-20 , Neoplasm Micrometastasis , Peritoneum , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger , Stomach Neoplasms
7.
Journal of the Korean Gastric Cancer Association ; : 128-133, 2003.
Article in Korean | WPRIM | ID: wpr-132067

ABSTRACT

PURPOSE: The benefits of the "no-touch" isolation technique that is usually performed to prevent the circulation of tumor cells are not evident. The aim of this study was to determine whether the no-touch isolation technique for treating gastrointestinal cancers could prevent the circulation of tumor cells detected by reverse transcriptase polymerase chain reaction (RT-PCR). Matrials and Methods: By using RT-PCR to amplify mRNAs for two specific epithelial markers, carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20), we examined 34 gastric cancer patients who had been histologically diagnosed and 22 patients had undergone serosal and peritoneal brushing. RESULTS: In 10 (29.4%) of the 34 gastric cancer patients, we detected CK20 mRNA before manipulation, and in 17 (51.5%) of those patients, after we detected it. The density of the CK20 mRNA band was increased in 11 cases (33.3%) and the density was decreased in 2 cases (6.1%). In 16 (48.5%) of the 34 gastric cancer patients, we detected CEA mRNA before manipulation, and in 17 (51.5%) patients after we detected it. The density of the CEA mRNA band was increased in 8 cases (24.2%) and decreased in 3 cases (9.1%). CONCLUSION: These result suggest that the "no-touch isolation technique" might be useful when operating on advanced gastric cancer patients and that serosal or Douglas pouch brushing can be used to determine the status of micrometastasis.


Subject(s)
Humans , Carcinoembryonic Antigen , Douglas' Pouch , Gastrointestinal Neoplasms , Keratin-20 , Neoplasm Micrometastasis , Peritoneum , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger , Stomach Neoplasms
8.
Journal of the Korean Surgical Society ; : 14-19, 2003.
Article in Korean | WPRIM | ID: wpr-51808

ABSTRACT

PURPOSE: Integrins are cell surface proteins that anchor the cells to the extra-cellular matrix. It has recently been found that integrins are involved in proliferations, migration, differentiation and survival signal transduction. We studied the expression of integrins in normal and cancer tissue of Korean breast cancer patients, and investigated the relationship between integrin expression and the characteristics of breast cancer. METHODS: Normal and malignant breast tissues were taken from 25 breast cancer patients who were admitted to the Ajou University Hospital. Specimens were immediately preserved in a nitrogen tank at the time of the operation. Total RNA was extracted, and semi-quantitative reverse transcriptase polymerase chain reactions (RT-PCR) performed with PCR primers for integrin alpha1, alpha2, alpha5, and alphav, and integrin beta1, and beta3. The integrin expressions were compared between the normal and malignant tissues, and the expressions were analyzed in relation to tumor characteristics. RESULTS: Integrin alpha1, alpha5, alphav, beta1, and beta3 were significantly over-expressed in breast cancer tissue than in normal tissue. There was no difference in integrin alpha2 expression between the normal and cancer tissues. Integrin beta1 was over-expressed to a greater extent in lower histological grade carcinomas and to a lesser extent in high grade tumors. Hormonal receptor positive tumor tissue had more alphav, alpha5, and beta1 integrin expressions. There was no significant relationship between integrins and tumor size, lymph node meta-stasis, lymphovascular involvement, or c-erb-B2 expression. CONCLUSIONS: Integrins alpha1, alpha5, alphav and beta3 were over- expressed in malignant breast tissue to a greater extent than in normal tissue. However, studies on the localization of integrin expression in cancer tissue, and co-relations of integrin over-expressions, with survival and drug sensitivity, must be followed to evaluate the clinical value of integrin expression.


Subject(s)
Humans , Integrin beta1 , Breast Neoplasms , Breast , Integrin alpha1 , Integrin alpha2 , Integrins , Lymph Nodes , Membrane Proteins , Nitrogen , Polymerase Chain Reaction , RNA , RNA-Directed DNA Polymerase , Signal Transduction
9.
Korean Journal of Obstetrics and Gynecology ; : 1736-1745, 2002.
Article in Korean | WPRIM | ID: wpr-37867

ABSTRACT

OBJECTIVE: The present study examined the gonadotropin regulation of TR3 gene expression by luteinizing hormone (LH) in cultured human luteinized granulosa cells. METHODS: TR3 mRNA levels were detected by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) method in cultured human luteinized granulosa cells collected from patients undergoing in vitro fertilization. RESULTS: TR3 transcript was transiently induced by LH, reaching maximum levels 1 hr after stimulation, in a dose-dependent manner. LH-stimulated TR3 expression was abolished by actinomycin D, but was superinduced by cycloheximide. Treatment of luteinized granulosa cells with Rp-cAMP, an inhibitor of protein kinase A, as well as, chelerythrin, an inhibitor of protein kinase C, suppressed LH-stimulated TR3 mRNA levels. In addition, forskolin and TPA mimicked the LH action on the induction of TR3 gene, implying the role of protein kinase A and C activation. CONCLUSION: Taken together, the present study demonstrates that TR3 gene was rapidly and transiently induced by LH in human luteinized granulosa cells. The results imply that TR3 may play a role in ovulation by initiating a cascade of ovulation-specific gene expression in response to LH.


Subject(s)
Female , Humans , Colforsin , Cyclic AMP-Dependent Protein Kinases , Cycloheximide , Dactinomycin , Fertilization in Vitro , Gene Expression , Gonadotropins , Granulosa Cells , Lutein , Luteinizing Hormone , Ovulation , Protein Kinase C , Receptors, Thyroid Hormone , RNA, Messenger , Thyroid Gland
10.
Korean Journal of Otolaryngology - Head and Neck Surgery ; : 963-968, 2002.
Article in Korean | WPRIM | ID: wpr-645409

ABSTRACT

BACKGROUND AND OBJECTIVES: Collectins (surfactant protein A and D) are proteins with collagen tails and globular lectin domains that appear to play an important role in the first line of host defense in mammalians. However, it is not known if collectins are also present in human nasal mucosa. The purpose of this study was to investigate the expression of collectin proteins in human nasal mucosa and to compare the expressions of SP-A and D mRNA in the normal nasal mucosa and in chronic inflammatory nasal diseases. MATERIALS AND METHOD: Ten chronic rhinosinusitis patients were recruited and ten normal nasal mucosae were used as normal controls. Reverse transcriptase-polymerase chain reaction was performed to detect SP-A and SP-D mRNA. The level of collectin and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) transcripts were semi-quantified with the desitometry. We have characterized the cellular localization of SP-A and SP-D protein using immunohistochemistry. RESULTS: SP-A2/GAPDH mRNA ratio in chronic rhinitis nasal mucosa is greater compared with that in normal nasal mucosa (p<0.05). SP-A protein was expressed in the nasal epithelium and in the epithelial cells of the submucosal glands. SP-D mRNA and protein were not expressed in the nasal mucosa. CONCLUSION: These data provide the first evidence of the presence of collectins in the human nasal mucosa. These results suggested that up-regulation of collectin in chronic rhinosinusitis may be a protective response for the nasal mucosa.


Subject(s)
Humans , Collagen , Collectins , Epithelial Cells , Immunohistochemistry , Nasal Mucosa , Nose Diseases , Pulmonary Surfactant-Associated Protein A , Pulmonary Surfactant-Associated Protein D , Pulmonary Surfactants , Rhinitis , RNA, Messenger , Sinusitis , Staphylococcal Protein A , Up-Regulation
11.
Chinese Journal of Medical Aesthetics and Cosmetology ; (6)2001.
Article in Chinese | WPRIM | ID: wpr-539955

ABSTRACT

Objective To evaluate the relative quantities of flt-1, KDR, endostatin mRNA in tissue specimens of human hemangioma and great saphfenous vein with the modified reverse transcriptase-polymerase chain reaction (RT-PCR). Methods Total RNA from human specimens by Trizol was reverse-transcribed and amplified by PCR in 4 deffirent tubes containing one of the primer pairs such as flt-1, KDR, endostatin or ?-actin. The target genes and beta-actin PCR products were about 200 bp in length. The ratio of the yield of the target gene PCR product to the beta-actin PCR product could be calculated after 35 cycles of amplification.Results These ratios were correlated positively ( P=0.014,P=0.019) in two groups, but no relationship in endostatine level was observed (P=0.436).Conclusions VEGFR may play a very important role in the pathogenesis of the congenital hemangioma in children. But the endostatin may take little effect on it.

12.
Journal of the Korean Surgical Society ; : 157-166, 1999.
Article in Korean | WPRIM | ID: wpr-45476

ABSTRACT

BACKGROUND: A high incidence of chronic liver disease is reported in end-stage renal failure patients due to hemodialysis and blood transfusion. An average of 20% of the patients who received renal hemodialysis are infected with hepatitis C virus, but the incidence of infection in these patients varies widely according to geographic location and the diagnostic methods used. Controversy exists regarding the impact of pretransplantation HCV infection on the outcome of renal transplantation. We measured the seroprevalence of the antibody to hepatitis C (anti-HCV) in renal transplant candidates and compared the prevalence of posttransplantation liver disease, graft, and patient survival among renal transplant recipients with and without anti-HCV at the time of the transplantation, and we attempted to define the possible factors affecting the clinical course following renal transplant in positive HCV patients. METHODS: Between June 1990 and December 1997, 634 patients underwent renal transplants at our institute. Viral infection with hepatitis were analyzed in these patients by using anti-HCV positivity using first, second, and third generation EIA, and RT-PCR. RESULTS: Twelve (12) of the 634 (1.9%) had positive anti-HCV before renal transplantation. During a mean follow-up of 29.4 months, viral mRNA was detected in the pretransplantation serum in 3 out of 8 (37.5%) positive anti-HCV patients. Among the 12 patients with positive anti-HCV, 2 (16.6%) showed early liver dysfunction, and 1 (8.3%) showed histologic progression to chronic hepatitis leading to hepatic failure and death. Graft loss occurred in 1 of the 12 (8.3%) patients with positive anti-HCV and in 62 of the 622 (9.8%) patients with negative anti-HCV. Three (3) out of the 12 (25%) patients with positive anti-HCV, and 121 of the 622 (19.6%) patients with negative anti-HCV had episodes of rejection. One (1) of the 12 (8.3%) patients with positive anti-HCV and 26 of the 622 (4.2%) patients with negative anti-HCV died after kidney transplantation. There were no statistical differences in patients or graft survival between the positive anti-HCV (+) and the negative anti-HCV patients. CONCLUSION: From these results, we can assume that the presence of anti-HCV without advance liver disease should not be a contraindication for kidney transplantation.


Subject(s)
Humans , Blood Transfusion , Follow-Up Studies , Graft Survival , Hepacivirus , Hepatitis , Hepatitis C , Hepatitis, Chronic , Incidence , Kidney Failure, Chronic , Kidney Transplantation , Liver Diseases , Liver Failure , Prevalence , Renal Dialysis , RNA, Messenger , Seroepidemiologic Studies , Transplantation , Transplants
13.
Korean Journal of Nephrology ; : 456-468, 1997.
Article in English | WPRIM | ID: wpr-151558

ABSTRACT

BACKGROUND: Korean hemorrhagic fever(KHF), a severe from of hemorrhagic fever with renal syndrome(HFRS), is the most common cause of acute renal failure in far east. Two serotypes of hantavirus, Hantaan and Seoul viruses, were identified as pathogens for KHF in Korea. To elucidate the diagnostic applicability for the serotype diagnosis in KHF patients, using a nested reverse transcriptase-PCR and restriction fragment length polymorphism(nRT-PCR /RFLP), we screened 4 prototype viruses, 11 virus isolates from KHF patients, and 69 specimens obtained from 30 KHF patients. METHODS: The nRT-PCR was performed using serotype specific primers for G1 segments for Hantaan(HF3 1140-1163, HB14 1363-1342) and Seoul(SF2 809-832, SB3 1200-1177) viruses. The PCR product was further amplified using nested primers for Hantaan(HF4, 1141-1164, HB13, 1360-1339) and Seoul(SF7 863-884, SB1 1165-1142) viruses. Amplified segments were digested with restriction enzymes specific for either Hantaan(Cla I) or Seoul(Sac I) virus sequences. In all cultured viruses, serotypes identified by nRT-PCR/RFLP were consistent with those of PRNT. RESULTS: In KHF patients, nRT-PCR/RFLP results were compatible with Hantaan virus in 10 patients and with Seoul virus in 13 patients. In 3 patients both Hantaan and Seoul specific amplified bands were visualized in serially collected samples, and in 4 patients no detectable amplicons were detected. Among 69 specimens, 55 specimens obtained from 3 to 33 day of illness were positive. The positive rate was affected by the hospital where specimens were collected, but not by clinical phases, the day of illness, or severity of HFRS. CONCLUSIONS: In general, nRT-PCR/RFLP was a rapid and convenient method for serotype diagnosis in most of the KHF patients. The presented method also make it possible to detect genetic variation of hantavirus within the same serotype. But unlike the viruses in culture, in testing patients' sera, the sensitivity of this methods needed to be improved especially by adequate sample handling.


Subject(s)
Humans , Acute Kidney Injury , Diagnosis , Asia, Eastern , Fever , Genetic Variation , Hantaan virus , Orthohantavirus , Hemorrhagic Fever with Renal Syndrome , Korea , Polymerase Chain Reaction , Seoul virus , Seoul
14.
The Journal of the Korean Rheumatism Association ; : 11-26, 1997.
Article in Korean | WPRIM | ID: wpr-55964

ABSTRACT

OBJECTIVE: To understand T cell role in the immunopathogenesis of rheumatoid arthritis(RA), authors investigated Vbeta usage of T cell receptor(TCR) in different onset RA lesion of the same patient using a reverse transcriptase-polymerase chain reaction. The current pathogenic model of RA plays a critical role in CD4+ T cells, which are thought to be able to recognize a disease-relevant antigen in the joint. In this model, activation of a certain, specific, antigen induced T cells plays a pivotal role in the development and maintenance of chronic inflammation. To search a common clone in different onset of inflammatory joints will furnish the most exact information about T cells which play a role at initiation and perpetuation of synovitis. Here, We first characterized the change in TCR Vbeta shape with elapsing time between the two joints that have the consecutive inflammation . METHODS: Synovial fluids and peripheral blood were obtained from a patient with active RA who had two successively developing inflammatory joints with 1 week interval(the right knee joint(RT) was involved first and the left knee joint(LT) later). RT-PCR technology was employed to examine synovial fluid and peripheral T cells. Oligonucleotide primers specific for individual TCR Vbeta gene families were used to amplify the TCR gene products in a semiquantitative assay of their relative utilization in fresh T cells subpopulations(CD4+, CD8+ T cells). RESULTS: The CD4+ T cells with TCR Vbeta 1(LT: 4.78%, RT: 2.17%), Vbeta 2(LT: 5.84%, RT: 0. 63%), Vbeta 5.1 (LT: 3.40%, RT: 1. 07%), Vbeta 5. 2 (LT: l. 91%, RT: 0. 31%), Vbeta 8(LT: 4. 20%,RT: 0. 19%), Vbeta 9(LT: 2. 61%, RT: 0. 12beta), Vbeta 10(LT: 4. 08%, RT: l. 27%), Vbeta 12(LT: 4. 53%, RT: 0. 99%), Vbeta 22 (LT: 3. 85%, RT: 0. 38%), Vbeta 23(LT: 3. 99%, RT: 0. 63%) were used more predominantly in the left knee joint than in the right knee joint while Vbeta 15 and 18 were used far more in the right knee joint. The CD8+ T cells were used less frequently in the left side than in the right side except the Vbeta 3, 4 and 7 families. CONCLUSIONS: Among the CD4+ T cells, TCR Vbeta 1, 2, 5. 1, 5. 2, 8, 9, 10, 12, 22, 23 families might play a key role in early symptomatic synovitis in RA. The role of TCR Vbeta 15 and 18 families increase in progressing synovitis with time. On the other hand, the late recruitment of CD8+ T cells in the inflamed site might be related to nonspecific inflammatory reaction.


Subject(s)
Humans , Arthritis, Rheumatoid , Clone Cells , DNA Primers , Genes, T-Cell Receptor , Hand , Inflammation , Joints , Knee Joint , Knee , Synovial Fluid , Synovitis , T-Lymphocytes
16.
Yonsei Medical Journal ; : 117-125, 1993.
Article in English | WPRIM | ID: wpr-37573

ABSTRACT

The 5'- and 3'-side half of liver type glucose transporter (GLUT2) cDNA was amplified from total RNA or mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). The amplified 5'-side fragment of GLUT2 cDNA was inserted into pGEM4Z and named pGLGT1, and the 3'-side fragment of GLUT2 cDNA was inserted into the HindIII site of pGLGT1 to construct pGLGT2 which contains an entire open reading frame of GLUT2 cDNA. The GLUT2 cDNA in pGLGT2 was transferred to an eukaryotic expression vector (pMAM) to construct pMLGT, which was expressed in the insulin-sensitive Chinese hamster ovary (CHO) cells. Western blot analysis showed that the GLUT2 gene in pMLGT was expressed in the transfected CHO cells successfully. The GLUT2 content in the plasma membrane fraction of insulin-treated CHO cells expressing GLUT2 increased 3.8-fold compared to that of the control group. This result suggests that GLUT2, which is not subjected to translocation by insulin in the cells of its major distribution, can be translocated if it is expressed in the suitable cells sensitive to insulin action.


Subject(s)
Animals , Base Sequence , CHO Cells , Cloning, Molecular , Cricetinae , Insulin/pharmacology , Liver/metabolism , Molecular Sequence Data , Monosaccharide Transport Proteins/genetics , Oligonucleotide Probes/genetics , Translocation, Genetic
17.
Chinese Journal of Blood Transfusion ; (12)1988.
Article in Chinese | WPRIM | ID: wpr-584739

ABSTRACT

Objective To evaluate the significance of HCV-RNA testing of blood donors by using reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and investigate HCV “window period” in blood donors.Methods Test blood donors for HCV-RNA by RT-PCR, and some positive cases were followed up. Results Among 28098 routine test negative blood donors, 77 (0.27%) were HCV-RNA positive. Eleven of the 77 HCV-RNA positive donors were followed up and 9 of them were later found to have increased ALT and/or positive anti-HCV. The interval between positive HCV-RNA test and abnormal serological tests was at least 4 weeks.Conclusion RT-PCR can identify HCV infected donors at an early stage. This test should become a routine test for blood donors to further ensure blood safety.

18.
Journal of Chongqing Medical University ; (12)1986.
Article in Chinese | WPRIM | ID: wpr-579391

ABSTRACT

Objective:To investigate the differential expression of VEGF165b in the renal cell carcinoma (RCC) and normal renal tissues and its role in the development of RCC. Methods: S-P immunohistochemistry was used to detect the expression of VEGF165b protein in 30 specimens of paraffin-embedded RCC tissues and 29 specimens of paraffin-embedded normal renal tissues. RT-PCR was performed to detect the expression of VEGF165b mRNA in 32 specimens of fresh RCC tissues and 30 specimens of fresh normal renal tissues. Results: Among 29 of normal renal tissues,28 specimens had positive expression of VEGF165b protein,with the positive rate of 96.55%(28/29) that was significantly higher than 20.00%(6/30)in RCC tissues(P

19.
Medical Journal of Chinese People's Liberation Army ; (12)1981.
Article in Chinese | WPRIM | ID: wpr-563575

ABSTRACT

Objective To study Axin expression in colorectal adenocarcinoma, colorectal adenoma, Peutz-Jeghers syndrome (PJS) polyp, and normal intestinal mucosa at mRNA and protein levels, and to explore the role of Axin in the pathogenesis of PJS. Methods The experiment samples, which were from the exsected normal mucosa, PJS polyps, adenomatous polyps and colorectal adenocarcinoma between Jan, 2001 and Mar, 2008 in Nanfang Hospital, each includes 32 sapamples. Immunohistochemistry was adopted to detect the Axin protein expression in each sample distinctively. Results transcriptase -polymerse chain reaction (RT-PCR) was used to detected and compare the Axin mRNA expression in 16 samples of each group. Results Of the tissues in each group, the positive Axin protein expression was located in the endochylema. The nomal mucosa group showed the most intensive expressive intensity, the olorectalc adenocarcinoma group was the least intensive, with significant difference in each group(P

20.
Acta Anatomica Sinica ; (6)1954.
Article in Chinese | WPRIM | ID: wpr-577937

ABSTRACT

Objective To compare the expression of pancreatic triglyceride lipase in type-1(T1A) and type-2 astrocytes(T2A).Methods Purified cultures of cortical T1A and T2A were prepared from neonatal rats.The expression of pancreatic triglyceride lipase mRNA was detected by reverse transcriptase polymerase chain reaction(RT-PCR).And the protein level was determined by immunocytochemistry with laser scanning confocal microscope.Results RT-PCR showed that the expression of pancreatic triglyceride lipase mRNA was negative in T1A and positive at a high level in T2A.Immunocytochemistry demonstrated that pancreatic triglyceride lipase expressed at a low level in T1A and was mainly distributed in the nuclei.By contrast,pancreatic triglyceride lipase expressed at a high level in T2A and was distributed diffusedly in the cytoplasm,nuclei and processes.Conclusion T1A and T2A showed different expression levels of pancreatic triglyceride lipase. Pancreatic triglyceride lipase expressed at a high level in T2A,which indicated that T2A may play an important role in the metabolism of lipids in the central nervous system.

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