Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR ...Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR and was cloned into pGEX-4T-1, a prokaryotic expressing vector. The recombinant plasmid containing rat α-Syn gene was transformed into E. Coli BL21 to express a fusion protein with rat α-Syn protein tagged by glutathione-S-transferase (GST). The fusion protein was then cleaved by thrombin during passing through the GST-agarose 4B column to release the recombinant rat α-Syn protein. The recombinant rat a-Syn protein was further purified using Superdex S200 gel filtration. Results DNA sequencing confirmed that the cloned cDNA contained 420 base pairs encoding 140 amino acids, which was identical to the reported amino acid sequence of rat α-Syn. After transformation, the recombinant plasmid pGEX-ra-Syn expressed a soluble protein that was inducible by IPTG. The purified recombinant protein was shown to be single band on SDS-PAGE, with a molecular size of around 18000, which was identical to the reported molecular size of rat α-Syn. Western blot analysis demonstrated that the recombinant protein was recognized by specific antibody against α-Syn. Conclusion The rat α-Syn gene was successfully expressed in prokaryotic expression system and highly purified rat α-Syn recombinant protein was produced.展开更多
AIM: To investigate the value of interleukin-8 (IL-8), a pro-inflammatory chemokine, in predicting the prognosis of pancreatic cancer. METHODS: Expression of IL-8 and its receptor CXCR1 was assessed by immunohistochem...AIM: To investigate the value of interleukin-8 (IL-8), a pro-inflammatory chemokine, in predicting the prognosis of pancreatic cancer. METHODS: Expression of IL-8 and its receptor CXCR1 was assessed by immunohistochemistry in pancreatic cancer and chronic pancreatitis samples. Enzyme-linked immunosorbent assay was used to detect the serum IL-8 levels in pancreatic cancer patients. Human pancreatic cancer tissues were heterotopically transplanted to the immune-deficiency mice to evaluate the effect of serum IL-8 on the tumorigenesis of the cancer samples.RESULTS: IL-8 and CXCR1 proteins were both overexpressed in pancreatic adenocarcinoma samples (55.6% and 65.4%, respectively) compared with the matched para-cancer tissues (25.9% and 12.3%, P < 0.01), or chronic pancreatitis (0% and 25%, P < 0.05). Serum IL-8 levels in pancreatic cancer patients (271.1 ± 187.7 ng/mL) were higher than in other digestive system tumors, such as gastric cancer (41.77 ± 9.11 ng/mL, P = 0.025), colorectal carcinoma (78.72 ± 80.60 ng/mL, P = 0.032) and hepatocellular carcinoma (59.60 ± 19.80 ng/mL, P = 0.016). In vivo tumorigenesis analysis further proved that tumor tissues from patients with higher serum IL-8 levels grew faster than those with lower IL-8 levels. CONCLUSION: IL-8 can be a fine serum marker for predicting the prognosis pancreatic cancer.展开更多
Objective: To investigate the expression and significance of cancer inhibitory gene PTEN protein in osteosar-coma. To analyze the level of its expression in different histological classification of osteosarcoma. To d...Objective: To investigate the expression and significance of cancer inhibitory gene PTEN protein in osteosar-coma. To analyze the level of its expression in different histological classification of osteosarcoma. To determine the possibility of taking PTEN protein as a marker gene for diagnosing osteosarcoma. To observe the clinical value of PTEN expression levels as a reference index for osteosarcoma classification. Methods: 43 specimens collected from osteosarcoma excision were studied. 30 specimens collected during the same period from benign lesion of bone (osteochondroma) operation were taken as the control group. Immunohistochemistry staining (ElivisonTM two steps method) was used to detect the expression of PTEN protein in 43 cases of osteosarcoma. SPSS 10.0 was used in statistical analysis. Results: Immunohistochemistry staining showed that the positive reaction of PTEN protein was all oriented to cytoplasm, which were brown or yellowish- brown granules. By way of X^2 test, the significant difference of the positive expressions of PTEN protein between bone benign lesion and osteosarcoma (X^2 = 7.976, P 〈 0.01) was observed. Osteosarcoma with different degrees of histodifferentiation showed different level expression of PTEN protein. There was significant difference between well-differentiated osteosarcoma (grades Ⅰ-Ⅱ) and poorly-differentiated osteosarcoma (grade Ⅲ) statistically (P 〈 0.01). The level of expression of PTEN was negatively correlated to the histological grade of osteosarcoma. There was great significance statistically (rs=-0.4922, P 〈 0.01). Conclusion: PTEN protein may be used as candidate gene of cancer inhibitory gene: PTEN protein is a cancer suppressor gene protein which has expression in bone tumors. It might not only be used in the study of pulmonary carcinoma and neurogliocytoma, but also in the study of bone tumor; the expression of PTEN is related to benignancy or malignancy of bone tumor and their degree of differentiation. The expression of PTEN is positively correlated with degree of differentiation.展开更多
Objective To examine the expression of cell division cycle associated 2(CDCA 2) in pancreatic ductal adenocarcinoma(PDAC) and investigate its role in prognosis of PDAC patients.Methods This retrospective study include...Objective To examine the expression of cell division cycle associated 2(CDCA 2) in pancreatic ductal adenocarcinoma(PDAC) and investigate its role in prognosis of PDAC patients.Methods This retrospective study included 155 PDAC patients who underwent surgical treatment and complete post-operative follow-up.Clinicopathologic data were collected through clinical database.Tissue microarray was constructed and immunohistochemistry was performed to detect CDCA2 expression in the PDAC tumor tissues and adjacent non-tumor tissues.Clinicopathological characteristics between high and low CDCA2 expression were compared.Correlation of CDCA2 expressions with patients' survival was analyzed using Kaplan-Meier method and Cox regression analysis.Results Expression of CDCA2 in PDAC cells was significantly higher than that in adjacent non-tumor tissues(U=4056.5,P<0.001).Univariate analysis showed that CDCA2 expression [hazard ratio(HR)=1.574,95% confidence interval(CI)=1.014-2.443,P=0.043] and node metastasis(HR=1.704,95%CI=1.183-2.454,P=0.004) were significantly associated with prognosis.Cox regression analysis showed CDCA2 expression was not an independent prognostic risk factor(HR=1.418,95%CI=0.897-2.242,P=0.135) for PDCA patients.Stratification survival analysis demonstrated CDCA2 expression as an independent prognostic risk factor in male patients(HR=2.554,95%CI=1.446-4.511,P=0.003) or in non-perineural invasion patients(HR=2.290,95%CI=1.146-4.577,P=0.012).Conclusions CDCA2 is highly expressed in PDAC tumor tissue.Although CDCA2 is not an independent prognostic risk factor for PDAC patients,it might be used to help predict prognosis of male or non-perineural invasion patients of PDAC.展开更多
Fish glue is the collagen from scale skin and bone of fish. It is known for value added product from fish processing and the adhesive agent for wood paper book binding etc. This work was aimed with the method to produ...Fish glue is the collagen from scale skin and bone of fish. It is known for value added product from fish processing and the adhesive agent for wood paper book binding etc. This work was aimed with the method to produce fish glue from fish skin through representing optimum fish type and its glue character by study on standard method for fish glue processing (using Oreochromis niloticus skin as the model), optimum fish type and characterization of the glue from optimum fish skin. Soaking fish skin in 0.1 N NaOH for 6 h and changed it every 3 h for pretreatment before acid extraction with 0.5 M acetic acid was the optimum condition for the standard processing. Among 20 types (O. niloticus, Oreochromis sp., Nemipterus sp., Psettodes erumei, Epiephelus malabaricus, Sphyraena obstsata, Channa striata, Xenentodon cancila, Barbonymus gonionotus, Liza vaigiensis, Anabas testudineus, Chanos chanos, Micronema bleekeri, Thunnus tonggol, Rastrelliger brachysom, Epinephelus lanceolatus, Lutjanus lineolatus, Pomadasys hasta, Selar crumenophthalmus and Sardaorentalis sp.), Chanos chanos was an appropriate type for glue production. Fat, protein, carbohydrate, moisture, pH and viscosity of Chanos chanos glue were 0.32%, 4.23%, 83.8%, 11.56%, 3.35, 4,978.33-8,180 cp, respectively. The glue was collagen type I which was composed ofal (148 kDa) and a2 (129 kDa) chain and could bind paper, wood and foam sheet.展开更多
AIM:To determine the role of CD133 in cholangiocarcinoma progression. METHODS:CD133 protein expression was evaluated by immunohistochemistry in 34 cholangiocarcinoma specimens.In addition,proliferation,chemoresistance...AIM:To determine the role of CD133 in cholangiocarcinoma progression. METHODS:CD133 protein expression was evaluated by immunohistochemistry in 34 cholangiocarcinoma specimens.In addition,proliferation,chemoresistance and invasive properties of CD133-enriched(CD133 + ) and CD133-depleted(CD133 )RMCCA1 cholangiocarcinoma cells were studied and compared. RESULTS:Strong CD133 expression was observed in 67.6%(23/34)of the cholangiocarcinoma specimens. Strong expression of CD133 was significantly associated with nodal metastasis(P=0.009)and positive surgical margin status(P=0.011).In the in vitro study, both the CD133 + and CD133 cells had similar proliferation abilities and resistance to chemotherapeutic drugs.However,the CD133 + cells had a higher invasive ability compared with CD133 cells. CONCLUSION:CD133+cells play an important role in the invasiveness of cholangiocarcinoma.Targeting of the CD133+cells may be a useful approach to improve treatment against cholangiocarcinoma.展开更多
AIM:To evaluate the usefulness of differentially expressed proteins from colorectal cancer (CRC) tissues for differentiating cancer and normal tissues.METHODS:A Proteomic approach was used to identify the differential...AIM:To evaluate the usefulness of differentially expressed proteins from colorectal cancer (CRC) tissues for differentiating cancer and normal tissues.METHODS:A Proteomic approach was used to identify the differentially expressed proteins between CRC and normal tissues.The proteins were extracted using Tris buffer and thiourea lysis buffer (TLB) for extraction of aqueous soluble and membrane-associated proteins,respectively.Chemometrics,namely principal component analysis (PCA) and linear discriminant analysis (LDA),were used to assess the usefulness of these proteins for identifying the cancerous state of tissues.RESULTS:Differentially expressed proteins identified were 37 aqueous soluble proteins in Tris extracts and 24 membrane-associated proteins in TLB extracts.Based on the protein spots intensity on 2D-gel images,PCA by applying an eigenvalue > 1 was successfully used to reduce the number of principal components (PCs) into 12 and seven PCs for Tris and TLB extracts,respectively,and subsequently six PCs,respectively from both the extracts were used for LDA.The LDA classification for Tris extract showed 82.7% of original samples were correctly classified,whereas 82.7% were correctly classified for the cross-validated samples.The LDA for TLB extract showed that 78.8% of original samples and 71.2% of the cross-validated samples were correctly classified.CONCLUSION:The classification of CRC tissues by PCA and LDA provided a promising distinction between normal and cancer types.These methods can possibly be used for identification of potential biomarkers among the differentially expressed proteins identified.展开更多
Over the few last years, cucumber (Cucumis sativus L) root rot disease became common and inflicted marked losses to yield in Fayoum. Isolation trails from infected cucumber roots revealed that Thielaviopsis basicola...Over the few last years, cucumber (Cucumis sativus L) root rot disease became common and inflicted marked losses to yield in Fayoum. Isolation trails from infected cucumber roots revealed that Thielaviopsis basicola and Fusarium moniliforme were the main cucumber root rot pathogens. The isolation trials from the rhizosphere of healthy cucumber plants revealed that two fungal isolates and four bacterial isolates had antagonistic effects against cucumber root rot fungi. All the tested biocontrol agents reduced the radial growth of all the tested root rots fungi in dual cultures. However, all culture filtrates of the tested biocontrol agent significantly reduced radial growth of all the tested pathogenic fungi, except that T. harzianum. Under field conditions, soil treatment with any of T. harzianum and B. subtilis BI and BF, significantly reduced percentages of infected plants and significantly increased percentages of survived plants and fruit yield compared with the control. Application of the commercial product Harpin protein (Messenger)~ product, as a resistance inducer at 0.3, 0.5 and l g/L significantly reduced the percentage of infected plants under greenhouse and field conditions. Field experiments indicated that the average percentage of infected plants after 90 days significantly decreased. The percentage of the survived plants as well as fruit yield increased by using integrated disease management (IDM) package, including the most effective treatments: tolerant cultivar, Trichoderma harzianum granules formula as soil treatment, Purging cassia plant extract, Harpin protein, and a half dose of Vitavax/thiram comparing with the treatment recommended by the Ministry of Agricultural or applied treatments individually.展开更多
GHB (γ-hydroxybutyrate) is becoming popular recreational drugs. As a result of its strong sedative and amnesiac effects, GHB has been implicated in a number of DFSA cases. The natural presence of GHB in the human b...GHB (γ-hydroxybutyrate) is becoming popular recreational drugs. As a result of its strong sedative and amnesiac effects, GHB has been implicated in a number of DFSA cases. The natural presence of GHB in the human body and its rapid elimination after ingestion make it difficult to detect and to evaluate its roles in suspected GHB-facilitated assaults. The paper describes an analytical method for the determination of GHB in urine using LC-MS/MS. Samples were acidified by ammonium chloride solution and extracted with ethyl acetate, and then the extracts were analysed by LC-MS/MS. The limit of detection was 0.05 p.g/mL (S/N = 3). The intra- and inter-day precision was within 10.0% at three concentrations. The methods were found to be sensitive, accurate, rapid and suitable for the forensic toxicology to test GHB in real cases.展开更多
Objective:The aim of the study was to investigate the expression of Smac protein in human hepatocarcinoma and their relationship with cell apoptosis and proliferation.Methods:The expressions of Smac and the proliferat...Objective:The aim of the study was to investigate the expression of Smac protein in human hepatocarcinoma and their relationship with cell apoptosis and proliferation.Methods:The expressions of Smac and the proliferating cell nuclear antigen (PCNA) in 41 cancer tissues,41 adjacent cirrhosis tissues and 9 normal control tissues in hemangioma were assessed by two-step immunohistochemical method and apoptosis was detected by TUNEL method.Results:Smac protein was expressed in 14 (34.14%) of the 41 cases of hepatocarcinoma,in 23 (56.10%) of the 41 cases of the adjacent cirrhosis tissues,and in 7 (77.8%) of the normal tissues in hemangioma.Smac protein positive expression rate in hepatocarcinoma was significantly lower than that in the adjacent cirrhosis tissues and the normal control tissues,χ2 were 3.989 and 4.115,respectively,and P were 0.046 and 0.042,respectively.Smac protein expression in cancer was significantly correlated with the ratio of apoptotic index to proliferative index,t'=2.260,P<0.05,but was not with the clinicopathological indicators such as the age and the histological grade,P>0.05.Conclusion:The relatively lower level of the expression of Smac may in a certain extent break the dynamic balance between apoptosis and proliferation of hepatocarcinoma cells,and then plays an important role in the pathogenesis of hepatocarcinoma.展开更多
Metallomics is proposed as a new omics to fol- low genomics, proteomics and metabolomics. This paper gives an overview of the development of met- allomics based on the introduction of the concept of metallomics and it...Metallomics is proposed as a new omics to fol- low genomics, proteomics and metabolomics. This paper gives an overview of the development of met- allomics based on the introduction of the concept of metallomics and its methodology.展开更多
Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media,...Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.展开更多
基金This work was supported by Key Project of National Natural Science Foundation of China (30430280) National Natural Science Foundation of China( 30271437,30270482 ) Natural Science Foundation of Beijing( 7022011 ).
文摘Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR and was cloned into pGEX-4T-1, a prokaryotic expressing vector. The recombinant plasmid containing rat α-Syn gene was transformed into E. Coli BL21 to express a fusion protein with rat α-Syn protein tagged by glutathione-S-transferase (GST). The fusion protein was then cleaved by thrombin during passing through the GST-agarose 4B column to release the recombinant rat α-Syn protein. The recombinant rat a-Syn protein was further purified using Superdex S200 gel filtration. Results DNA sequencing confirmed that the cloned cDNA contained 420 base pairs encoding 140 amino acids, which was identical to the reported amino acid sequence of rat α-Syn. After transformation, the recombinant plasmid pGEX-ra-Syn expressed a soluble protein that was inducible by IPTG. The purified recombinant protein was shown to be single band on SDS-PAGE, with a molecular size of around 18000, which was identical to the reported molecular size of rat α-Syn. Western blot analysis demonstrated that the recombinant protein was recognized by specific antibody against α-Syn. Conclusion The rat α-Syn gene was successfully expressed in prokaryotic expression system and highly purified rat α-Syn recombinant protein was produced.
基金Supported by The National Key Project of Scientific and Technical Supporting Programs of China, No. 2006BAI02A14National Natural Science Foundation of China, No. 30770996 and No. 30901776
文摘AIM: To investigate the value of interleukin-8 (IL-8), a pro-inflammatory chemokine, in predicting the prognosis of pancreatic cancer. METHODS: Expression of IL-8 and its receptor CXCR1 was assessed by immunohistochemistry in pancreatic cancer and chronic pancreatitis samples. Enzyme-linked immunosorbent assay was used to detect the serum IL-8 levels in pancreatic cancer patients. Human pancreatic cancer tissues were heterotopically transplanted to the immune-deficiency mice to evaluate the effect of serum IL-8 on the tumorigenesis of the cancer samples.RESULTS: IL-8 and CXCR1 proteins were both overexpressed in pancreatic adenocarcinoma samples (55.6% and 65.4%, respectively) compared with the matched para-cancer tissues (25.9% and 12.3%, P < 0.01), or chronic pancreatitis (0% and 25%, P < 0.05). Serum IL-8 levels in pancreatic cancer patients (271.1 ± 187.7 ng/mL) were higher than in other digestive system tumors, such as gastric cancer (41.77 ± 9.11 ng/mL, P = 0.025), colorectal carcinoma (78.72 ± 80.60 ng/mL, P = 0.032) and hepatocellular carcinoma (59.60 ± 19.80 ng/mL, P = 0.016). In vivo tumorigenesis analysis further proved that tumor tissues from patients with higher serum IL-8 levels grew faster than those with lower IL-8 levels. CONCLUSION: IL-8 can be a fine serum marker for predicting the prognosis pancreatic cancer.
文摘Objective: To investigate the expression and significance of cancer inhibitory gene PTEN protein in osteosar-coma. To analyze the level of its expression in different histological classification of osteosarcoma. To determine the possibility of taking PTEN protein as a marker gene for diagnosing osteosarcoma. To observe the clinical value of PTEN expression levels as a reference index for osteosarcoma classification. Methods: 43 specimens collected from osteosarcoma excision were studied. 30 specimens collected during the same period from benign lesion of bone (osteochondroma) operation were taken as the control group. Immunohistochemistry staining (ElivisonTM two steps method) was used to detect the expression of PTEN protein in 43 cases of osteosarcoma. SPSS 10.0 was used in statistical analysis. Results: Immunohistochemistry staining showed that the positive reaction of PTEN protein was all oriented to cytoplasm, which were brown or yellowish- brown granules. By way of X^2 test, the significant difference of the positive expressions of PTEN protein between bone benign lesion and osteosarcoma (X^2 = 7.976, P 〈 0.01) was observed. Osteosarcoma with different degrees of histodifferentiation showed different level expression of PTEN protein. There was significant difference between well-differentiated osteosarcoma (grades Ⅰ-Ⅱ) and poorly-differentiated osteosarcoma (grade Ⅲ) statistically (P 〈 0.01). The level of expression of PTEN was negatively correlated to the histological grade of osteosarcoma. There was great significance statistically (rs=-0.4922, P 〈 0.01). Conclusion: PTEN protein may be used as candidate gene of cancer inhibitory gene: PTEN protein is a cancer suppressor gene protein which has expression in bone tumors. It might not only be used in the study of pulmonary carcinoma and neurogliocytoma, but also in the study of bone tumor; the expression of PTEN is related to benignancy or malignancy of bone tumor and their degree of differentiation. The expression of PTEN is positively correlated with degree of differentiation.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(2012AA02A212)
文摘Objective To examine the expression of cell division cycle associated 2(CDCA 2) in pancreatic ductal adenocarcinoma(PDAC) and investigate its role in prognosis of PDAC patients.Methods This retrospective study included 155 PDAC patients who underwent surgical treatment and complete post-operative follow-up.Clinicopathologic data were collected through clinical database.Tissue microarray was constructed and immunohistochemistry was performed to detect CDCA2 expression in the PDAC tumor tissues and adjacent non-tumor tissues.Clinicopathological characteristics between high and low CDCA2 expression were compared.Correlation of CDCA2 expressions with patients' survival was analyzed using Kaplan-Meier method and Cox regression analysis.Results Expression of CDCA2 in PDAC cells was significantly higher than that in adjacent non-tumor tissues(U=4056.5,P<0.001).Univariate analysis showed that CDCA2 expression [hazard ratio(HR)=1.574,95% confidence interval(CI)=1.014-2.443,P=0.043] and node metastasis(HR=1.704,95%CI=1.183-2.454,P=0.004) were significantly associated with prognosis.Cox regression analysis showed CDCA2 expression was not an independent prognostic risk factor(HR=1.418,95%CI=0.897-2.242,P=0.135) for PDCA patients.Stratification survival analysis demonstrated CDCA2 expression as an independent prognostic risk factor in male patients(HR=2.554,95%CI=1.446-4.511,P=0.003) or in non-perineural invasion patients(HR=2.290,95%CI=1.146-4.577,P=0.012).Conclusions CDCA2 is highly expressed in PDAC tumor tissue.Although CDCA2 is not an independent prognostic risk factor for PDAC patients,it might be used to help predict prognosis of male or non-perineural invasion patients of PDAC.
文摘Fish glue is the collagen from scale skin and bone of fish. It is known for value added product from fish processing and the adhesive agent for wood paper book binding etc. This work was aimed with the method to produce fish glue from fish skin through representing optimum fish type and its glue character by study on standard method for fish glue processing (using Oreochromis niloticus skin as the model), optimum fish type and characterization of the glue from optimum fish skin. Soaking fish skin in 0.1 N NaOH for 6 h and changed it every 3 h for pretreatment before acid extraction with 0.5 M acetic acid was the optimum condition for the standard processing. Among 20 types (O. niloticus, Oreochromis sp., Nemipterus sp., Psettodes erumei, Epiephelus malabaricus, Sphyraena obstsata, Channa striata, Xenentodon cancila, Barbonymus gonionotus, Liza vaigiensis, Anabas testudineus, Chanos chanos, Micronema bleekeri, Thunnus tonggol, Rastrelliger brachysom, Epinephelus lanceolatus, Lutjanus lineolatus, Pomadasys hasta, Selar crumenophthalmus and Sardaorentalis sp.), Chanos chanos was an appropriate type for glue production. Fat, protein, carbohydrate, moisture, pH and viscosity of Chanos chanos glue were 0.32%, 4.23%, 83.8%, 11.56%, 3.35, 4,978.33-8,180 cp, respectively. The glue was collagen type I which was composed ofal (148 kDa) and a2 (129 kDa) chain and could bind paper, wood and foam sheet.
基金Supported by Rajavithi Hospital Project Grant and Thailand Research Fund(RSA52)
文摘AIM:To determine the role of CD133 in cholangiocarcinoma progression. METHODS:CD133 protein expression was evaluated by immunohistochemistry in 34 cholangiocarcinoma specimens.In addition,proliferation,chemoresistance and invasive properties of CD133-enriched(CD133 + ) and CD133-depleted(CD133 )RMCCA1 cholangiocarcinoma cells were studied and compared. RESULTS:Strong CD133 expression was observed in 67.6%(23/34)of the cholangiocarcinoma specimens. Strong expression of CD133 was significantly associated with nodal metastasis(P=0.009)and positive surgical margin status(P=0.011).In the in vitro study, both the CD133 + and CD133 cells had similar proliferation abilities and resistance to chemotherapeutic drugs.However,the CD133 + cells had a higher invasive ability compared with CD133 cells. CONCLUSION:CD133+cells play an important role in the invasiveness of cholangiocarcinoma.Targeting of the CD133+cells may be a useful approach to improve treatment against cholangiocarcinoma.
基金Supported by Research Universiti Grant,Grant No. 1001/PFAR MASI/815007
文摘AIM:To evaluate the usefulness of differentially expressed proteins from colorectal cancer (CRC) tissues for differentiating cancer and normal tissues.METHODS:A Proteomic approach was used to identify the differentially expressed proteins between CRC and normal tissues.The proteins were extracted using Tris buffer and thiourea lysis buffer (TLB) for extraction of aqueous soluble and membrane-associated proteins,respectively.Chemometrics,namely principal component analysis (PCA) and linear discriminant analysis (LDA),were used to assess the usefulness of these proteins for identifying the cancerous state of tissues.RESULTS:Differentially expressed proteins identified were 37 aqueous soluble proteins in Tris extracts and 24 membrane-associated proteins in TLB extracts.Based on the protein spots intensity on 2D-gel images,PCA by applying an eigenvalue > 1 was successfully used to reduce the number of principal components (PCs) into 12 and seven PCs for Tris and TLB extracts,respectively,and subsequently six PCs,respectively from both the extracts were used for LDA.The LDA classification for Tris extract showed 82.7% of original samples were correctly classified,whereas 82.7% were correctly classified for the cross-validated samples.The LDA for TLB extract showed that 78.8% of original samples and 71.2% of the cross-validated samples were correctly classified.CONCLUSION:The classification of CRC tissues by PCA and LDA provided a promising distinction between normal and cancer types.These methods can possibly be used for identification of potential biomarkers among the differentially expressed proteins identified.
文摘Over the few last years, cucumber (Cucumis sativus L) root rot disease became common and inflicted marked losses to yield in Fayoum. Isolation trails from infected cucumber roots revealed that Thielaviopsis basicola and Fusarium moniliforme were the main cucumber root rot pathogens. The isolation trials from the rhizosphere of healthy cucumber plants revealed that two fungal isolates and four bacterial isolates had antagonistic effects against cucumber root rot fungi. All the tested biocontrol agents reduced the radial growth of all the tested root rots fungi in dual cultures. However, all culture filtrates of the tested biocontrol agent significantly reduced radial growth of all the tested pathogenic fungi, except that T. harzianum. Under field conditions, soil treatment with any of T. harzianum and B. subtilis BI and BF, significantly reduced percentages of infected plants and significantly increased percentages of survived plants and fruit yield compared with the control. Application of the commercial product Harpin protein (Messenger)~ product, as a resistance inducer at 0.3, 0.5 and l g/L significantly reduced the percentage of infected plants under greenhouse and field conditions. Field experiments indicated that the average percentage of infected plants after 90 days significantly decreased. The percentage of the survived plants as well as fruit yield increased by using integrated disease management (IDM) package, including the most effective treatments: tolerant cultivar, Trichoderma harzianum granules formula as soil treatment, Purging cassia plant extract, Harpin protein, and a half dose of Vitavax/thiram comparing with the treatment recommended by the Ministry of Agricultural or applied treatments individually.
文摘GHB (γ-hydroxybutyrate) is becoming popular recreational drugs. As a result of its strong sedative and amnesiac effects, GHB has been implicated in a number of DFSA cases. The natural presence of GHB in the human body and its rapid elimination after ingestion make it difficult to detect and to evaluate its roles in suspected GHB-facilitated assaults. The paper describes an analytical method for the determination of GHB in urine using LC-MS/MS. Samples were acidified by ammonium chloride solution and extracted with ethyl acetate, and then the extracts were analysed by LC-MS/MS. The limit of detection was 0.05 p.g/mL (S/N = 3). The intra- and inter-day precision was within 10.0% at three concentrations. The methods were found to be sensitive, accurate, rapid and suitable for the forensic toxicology to test GHB in real cases.
文摘Objective:The aim of the study was to investigate the expression of Smac protein in human hepatocarcinoma and their relationship with cell apoptosis and proliferation.Methods:The expressions of Smac and the proliferating cell nuclear antigen (PCNA) in 41 cancer tissues,41 adjacent cirrhosis tissues and 9 normal control tissues in hemangioma were assessed by two-step immunohistochemical method and apoptosis was detected by TUNEL method.Results:Smac protein was expressed in 14 (34.14%) of the 41 cases of hepatocarcinoma,in 23 (56.10%) of the 41 cases of the adjacent cirrhosis tissues,and in 7 (77.8%) of the normal tissues in hemangioma.Smac protein positive expression rate in hepatocarcinoma was significantly lower than that in the adjacent cirrhosis tissues and the normal control tissues,χ2 were 3.989 and 4.115,respectively,and P were 0.046 and 0.042,respectively.Smac protein expression in cancer was significantly correlated with the ratio of apoptotic index to proliferative index,t'=2.260,P<0.05,but was not with the clinicopathological indicators such as the age and the histological grade,P>0.05.Conclusion:The relatively lower level of the expression of Smac may in a certain extent break the dynamic balance between apoptosis and proliferation of hepatocarcinoma cells,and then plays an important role in the pathogenesis of hepatocarcinoma.
基金supported by the National Basic Research Program of China(2003CB4 15001)the National Natural Science Foundation of China(20205008,20477053),
文摘Metallomics is proposed as a new omics to fol- low genomics, proteomics and metabolomics. This paper gives an overview of the development of met- allomics based on the introduction of the concept of metallomics and its methodology.
基金supported by the National Natural Science Foundation of China (Grant No. 209750240)the National Basic Research Program of China (Grant No. 2010CB912700)
文摘Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.