AIM: To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through ...AIM: To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through analyzing a variety of protein maps.METHODS: Two-DE profiles of HCT116 were established in IN-treated and untreated groups. Total proteins were separated by immobilized pH gradient-based 2-DE. The gels were stained by silver, scanned by ImageScanner,and analyzed with Image Master software.RESULTS: Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated group. The average deviation of spot position was 0.896±0.177 mm in IEF direction and 1.106±0.289 mm in SDS-PAGE direction respectively. In IN-treated group,1 169±36 spots were detected and 1 061±32 spots were matched, the average matching rate was 90.6% in three gels. In untreated group, 1 256±50 spots were detected and 1 168±46 spots were matched, the average matching rate was 93.0% in three gels. Forty-five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 protein spots decreased and 9showed higher expression in IN-treated group, and only two protein spots showed an expression in untreated cells.CONCLUSION: Two-DE profiles of IN-treated and untreated HCT116 cells were established. Apparent 45 different protein spots were detected in IN-treated and untreated HCT116 cells. The analysis on differential protein spots may serve as a new way to study the molecule mechanism of IN-treated colon cancer.展开更多
Objective.In 2003 we identified a family with familial hypocalciuric hypercalcemia(FHH)(heterozygous CASR gene mutation L173P)and a mutation in the pancreatic secretory trypsin inhibitor gene(SPINK1)(N34S).While famil...Objective.In 2003 we identified a family with familial hypocalciuric hypercalcemia(FHH)(heterozygous CASR gene mutation L173P)and a mutation in the pancreatic secretory trypsin inhibitor gene(SPINK1)(N34S).While family members with an isolated calcium-sensing receptor gene(CASR)mutation remained healthy,a combination of the CASR and SPINK1 gene mutation caused chronic pancreatitis(CP).We thus speculate that the combination of two genetic defects affecting calcium homeostasis and pancreatic enzyme activation might represent a novel approach in chronic inherited pancreatic disease.We therefore sought to explore whether CASR gene mutations were prevalent in a cohort of patients with CP and confirmed SPINK1 mutations.Material and methods.A cohort of 19 families(n = 170)with a history of idiopathic CP(ICP)was screened for mutations within the CASR gene;104 members of that cohort had a mutation(N34S)within the SPINK1 gene and 66 of those were suffering from CP.The entire CASR gene was screened for single strand conformation polymorphism under varying polyacrylamide gel conditions and subjected to direct dideoxy nucleotide sequencing of amplified cDNA.Results.Single-strand conformation polymorphisms were observed in 59 samples,clustering of exons 3,4 and 7.DNA sequence analysis revealed a yet unreported missense mutation in exon 7(R896H)and two conservative mutations in exon 4(F391F)and exon 7(E790E).Furthermore,an intronic polymorphism in nucleotide position 493-19 G > A was detected in 19 out of 170 members of that cohort.Conclusions.We identified three novel calcium-sensing receptor gene mutations(1 missense mutation,2 silent mutations and 1 intronic polymorphism)in a cohort of 19 families with ICP.In particular,the kindred with the R896H mutation presenting with a similar pedigree to the family described above may indicate a role for CASR gene mutations in SPINK1-related CP.Again,only the patient with the combination of both CASR and N34S SPINK1 gene mutation developed pancreatitis,whereas in the healthy parents and children only an isolated CASR or N34S SPINK1 gene mutation could be detected.We suggest that the CASR gene is a novel yet undetected co-factor in a multifactorial genetic setting of SPINK1-related pancreatitis that alters the susceptibility for pancreatitis in these patients.展开更多
AIM:To investigate the association between hepatocel-lular carcinoma (HCC) susceptibility and a 12-bp inser-tion/deletion polymorphism (rs6147150) in the 3'UTR of ErbB4.METHODS:Using a case-control design,the rs61...AIM:To investigate the association between hepatocel-lular carcinoma (HCC) susceptibility and a 12-bp inser-tion/deletion polymorphism (rs6147150) in the 3'UTR of ErbB4.METHODS:Using a case-control design,the rs6147150 genotypes in 270 patients with HCC and 270 healthy controls were determined by direct polymerase chain reaction and polyacrylamide gel electrophoresis.Logistic regression was used to analyze the association between the polymorphism and cancer risk.RESULTS:Computational modeling suggested that rs6147150 was located in the seed region of hsa-let-7c,a potential target sequence in ErbB4 3'UTR.Logistic re-gression analysis showed that,compared with individu-als homozygous for wild-type,heterozygotes [adjusted odds ratio (OR)=1.48,95% confidence interval (CI)= 1.03-2.17,P=0.034] and individuals homozygous for 12-bp del/del (OR=2.50,95% CI=1.37-4.56,P=0.001) were at significantly higher risk of HCC.Car-riers of the "del" allele of rs6147150 had a 1.59-fold increased risk for HCC (95% CI=1.22-2.07,P=0.003).CONCLUSION:rs6147150 may be associated with HCC risk,in part through let-7c-mediated regulation,and may be involved in the pathogenesis of HCC in Chi-nese populations.展开更多
A group of grafted PET fibers with different graft yield are formed by grafting acrylamide onto the PET main chains. The structure of grafted fibers are studied by scanning electronic microscope ( SEM ), infra-red spe...A group of grafted PET fibers with different graft yield are formed by grafting acrylamide onto the PET main chains. The structure of grafted fibers are studied by scanning electronic microscope ( SEM ), infra-red spectrophotometer ( IR ), and differential scanning calorimetry(DSC). At the same time, the moisture regain, dyeability, strength, and elongation at break of the samples are measured and their relations with structural changes are discussed. Compared with ungrafted fiber, shape of the fiber cross-section, IR characteristic absorption peaks, and melting behavior of the grafted fibers have been changed, causing the fiber dyeability and moisture regain to be increased, and mechanical properties to be changed.展开更多
AIM: To comprehensively identify the proteins of tumor relative antigen Ca-Hb3 recognized by colorectal carcinoma monoclonal antibody Hb3. METHODS: Ca-Hb3 was isolated by SDS-polyacrylamide gel electrophoresis (PAGE) ...AIM: To comprehensively identify the proteins of tumor relative antigen Ca-Hb3 recognized by colorectal carcinoma monoclonal antibody Hb3. METHODS: Ca-Hb3 was isolated by SDS-polyacrylamide gel electrophoresis (PAGE) followed by digestion with trypsin. Trypsin peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/ MS). The proteins identified by mass spectrometry were analyzed using bioinformatics. RESULTS: Ca-Hb3 was identified as a CKAP4-like protein by Nano HPLC tandem mass spectrometry analysis. The molecular weight of CKAP4-like protein was 62.02 kDa, including one hydrophobic region, one transmembrane domain, five coiled coils, four glycosylation sites and forty-nine phosphorylation sites. CKAP4-like protein had a high homogeneity with DeltaNp63α. The characteristic expression of DeltaNp63α that is considered a potential oncogene in the isoforms of p63 was similar to that of Ca-Hb3. CONCLUSION: Ca-Hb3 is probably a CKAP4-like protein, belonging to DeltaNp63α isoform of p63 family.展开更多
基金Supported by the National Natural Science Foundation of China, No. 30271516
文摘AIM: To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through analyzing a variety of protein maps.METHODS: Two-DE profiles of HCT116 were established in IN-treated and untreated groups. Total proteins were separated by immobilized pH gradient-based 2-DE. The gels were stained by silver, scanned by ImageScanner,and analyzed with Image Master software.RESULTS: Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated group. The average deviation of spot position was 0.896±0.177 mm in IEF direction and 1.106±0.289 mm in SDS-PAGE direction respectively. In IN-treated group,1 169±36 spots were detected and 1 061±32 spots were matched, the average matching rate was 90.6% in three gels. In untreated group, 1 256±50 spots were detected and 1 168±46 spots were matched, the average matching rate was 93.0% in three gels. Forty-five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 protein spots decreased and 9showed higher expression in IN-treated group, and only two protein spots showed an expression in untreated cells.CONCLUSION: Two-DE profiles of IN-treated and untreated HCT116 cells were established. Apparent 45 different protein spots were detected in IN-treated and untreated HCT116 cells. The analysis on differential protein spots may serve as a new way to study the molecule mechanism of IN-treated colon cancer.
文摘Objective.In 2003 we identified a family with familial hypocalciuric hypercalcemia(FHH)(heterozygous CASR gene mutation L173P)and a mutation in the pancreatic secretory trypsin inhibitor gene(SPINK1)(N34S).While family members with an isolated calcium-sensing receptor gene(CASR)mutation remained healthy,a combination of the CASR and SPINK1 gene mutation caused chronic pancreatitis(CP).We thus speculate that the combination of two genetic defects affecting calcium homeostasis and pancreatic enzyme activation might represent a novel approach in chronic inherited pancreatic disease.We therefore sought to explore whether CASR gene mutations were prevalent in a cohort of patients with CP and confirmed SPINK1 mutations.Material and methods.A cohort of 19 families(n = 170)with a history of idiopathic CP(ICP)was screened for mutations within the CASR gene;104 members of that cohort had a mutation(N34S)within the SPINK1 gene and 66 of those were suffering from CP.The entire CASR gene was screened for single strand conformation polymorphism under varying polyacrylamide gel conditions and subjected to direct dideoxy nucleotide sequencing of amplified cDNA.Results.Single-strand conformation polymorphisms were observed in 59 samples,clustering of exons 3,4 and 7.DNA sequence analysis revealed a yet unreported missense mutation in exon 7(R896H)and two conservative mutations in exon 4(F391F)and exon 7(E790E).Furthermore,an intronic polymorphism in nucleotide position 493-19 G > A was detected in 19 out of 170 members of that cohort.Conclusions.We identified three novel calcium-sensing receptor gene mutations(1 missense mutation,2 silent mutations and 1 intronic polymorphism)in a cohort of 19 families with ICP.In particular,the kindred with the R896H mutation presenting with a similar pedigree to the family described above may indicate a role for CASR gene mutations in SPINK1-related CP.Again,only the patient with the combination of both CASR and N34S SPINK1 gene mutation developed pancreatitis,whereas in the healthy parents and children only an isolated CASR or N34S SPINK1 gene mutation could be detected.We suggest that the CASR gene is a novel yet undetected co-factor in a multifactorial genetic setting of SPINK1-related pancreatitis that alters the susceptibility for pancreatitis in these patients.
基金Supported by The Applied Basic Research Programs of Science and Technology Commission Foundation of Suzhou,No.sys201047
文摘AIM:To investigate the association between hepatocel-lular carcinoma (HCC) susceptibility and a 12-bp inser-tion/deletion polymorphism (rs6147150) in the 3'UTR of ErbB4.METHODS:Using a case-control design,the rs6147150 genotypes in 270 patients with HCC and 270 healthy controls were determined by direct polymerase chain reaction and polyacrylamide gel electrophoresis.Logistic regression was used to analyze the association between the polymorphism and cancer risk.RESULTS:Computational modeling suggested that rs6147150 was located in the seed region of hsa-let-7c,a potential target sequence in ErbB4 3'UTR.Logistic re-gression analysis showed that,compared with individu-als homozygous for wild-type,heterozygotes [adjusted odds ratio (OR)=1.48,95% confidence interval (CI)= 1.03-2.17,P=0.034] and individuals homozygous for 12-bp del/del (OR=2.50,95% CI=1.37-4.56,P=0.001) were at significantly higher risk of HCC.Car-riers of the "del" allele of rs6147150 had a 1.59-fold increased risk for HCC (95% CI=1.22-2.07,P=0.003).CONCLUSION:rs6147150 may be associated with HCC risk,in part through let-7c-mediated regulation,and may be involved in the pathogenesis of HCC in Chi-nese populations.
文摘A group of grafted PET fibers with different graft yield are formed by grafting acrylamide onto the PET main chains. The structure of grafted fibers are studied by scanning electronic microscope ( SEM ), infra-red spectrophotometer ( IR ), and differential scanning calorimetry(DSC). At the same time, the moisture regain, dyeability, strength, and elongation at break of the samples are measured and their relations with structural changes are discussed. Compared with ungrafted fiber, shape of the fiber cross-section, IR characteristic absorption peaks, and melting behavior of the grafted fibers have been changed, causing the fiber dyeability and moisture regain to be increased, and mechanical properties to be changed.
基金The National Natural Science Foundation of China, No. 30300155
文摘AIM: To comprehensively identify the proteins of tumor relative antigen Ca-Hb3 recognized by colorectal carcinoma monoclonal antibody Hb3. METHODS: Ca-Hb3 was isolated by SDS-polyacrylamide gel electrophoresis (PAGE) followed by digestion with trypsin. Trypsin peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/ MS). The proteins identified by mass spectrometry were analyzed using bioinformatics. RESULTS: Ca-Hb3 was identified as a CKAP4-like protein by Nano HPLC tandem mass spectrometry analysis. The molecular weight of CKAP4-like protein was 62.02 kDa, including one hydrophobic region, one transmembrane domain, five coiled coils, four glycosylation sites and forty-nine phosphorylation sites. CKAP4-like protein had a high homogeneity with DeltaNp63α. The characteristic expression of DeltaNp63α that is considered a potential oncogene in the isoforms of p63 was similar to that of Ca-Hb3. CONCLUSION: Ca-Hb3 is probably a CKAP4-like protein, belonging to DeltaNp63α isoform of p63 family.
