OBJECTIVE To investigate the available parameters in gynecological screening for cervical lesions by liquid-based cytology technology （ThinPrep Cytology Test, TCT） and The Bethesda System （TBS）, also with computer...OBJECTIVE To investigate the available parameters in gynecological screening for cervical lesions by liquid-based cytology technology （ThinPrep Cytology Test, TCT） and The Bethesda System （TBS）, also with computer image analysis. METHODS With application of the image analysis system, all grades of cervical lesion cells were detected quantitatively and sorted in atypical squamous cells of undetermined significance （ASCUS）, atypical squamous cells-cannot exclude HSIL （ASC-H）, low-grade squamous intraepithelial lesion （LSIL）, high-grade squamous intraepithelial lesion （HSIL） and cervical squamous cell carcinoma （SCC） with the mean optical density （MOD）, average grey （AG）, positive units （PU）, and nucleus to cytoplasmic ratio （N： C）. Differences between each group of cells were compared and analyzed statistically. RESULTS Apart from four stereologic parameters in LSIL and HSIL groups there were no differences among them, in the other groups, there was statistically significant in differences between MOD, AG and PU values. Differences between them in the ratio of nucleus to cytoplasm were highly statistically significant. CONCLUSION Stereological indexes may serve as a screening tool for cervical lesions. The image analysis system is expected to become a new means of cytological assisted diagnosis.展开更多
In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the f...In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.展开更多
The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with...The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with which the Th cells status of the patients with gynaecological tumors and chronic renal failure （CRF） can be analyzed. IL-2 eDNA and IL-4 eDNA were prepared, and the plasmid pMD18 carrying IL-2 eDNA or IL-4 eDNA fragment was constructed and cloned as the template for quantitative determination. The primers and probes labelled with 6-carbexy-fluorescein （FAM） and 6-carboxy- tetramethylrhodamine （TAMRA） were prepared, and the experimental conditions were optimized to set up the FQ-RT-PCR method for quantitative determination of IL-2 mRNA and IL-4 mRNA. Th cells enriched from peripheral blood mononuclear cells （PBMCs） of 20 healthy volunteers （HVs）, 16 gynaecological benign （GB） cases, 18 gynaecological malignant （GM） tumor cases and 16 chronic renal failure （CRF） patients were tested for IL-2 mRNA and IL-4 mRNA by FQ-RT-PCR. The house-keeping gene β-actin was used as the internal control gene of the experiment. The standard curve for log concentration of series of quantitative templates vs threshold cycle （CT） was established by linear regression, and the linear range was 102-107 copies/μl. The imprecision test showed the CV of inter-assay and intra-assay of a high cont- ent sample by FQ-RT-PCR were 7.8% and 12.5%, respectively. The CV of inter-assay and intra-assay of a low content sample were 10.8% and 19.5%, respectively. The IL-2 mRNA expressions in Th of the patients with gynaecological malignant tumor （compared with the HVs and the patients with gynaecological benign disease） and in Th of the CRF patients （compared with the HVs） were declined significantly and at the same time the IL-4 mRNA expression increased significantly （ P 〈 0. 001 ）. A simple, sensitive and accurate FQ-RT-PCR method for the quantitative detection of IL-2 mRNA and IL-4 mRNA has been established. The IL-2 mRNA and IL-4 mRNA expressions in Th cells of the patients with gynaecological malignant tumor and the CRF patients were polarized and displayed Th2 response. It suggests that the function of the Th cells of the patients with gynaecological malignant tumor or CRF is at unbalance.展开更多
A novel sensitive semi-quantitative virus detection technique was developed using the respiratory syncytial virus(RSV) as an example, through dark-field light scattering imaging of the surface state of the virusinvade...A novel sensitive semi-quantitative virus detection technique was developed using the respiratory syncytial virus(RSV) as an example, through dark-field light scattering imaging of the surface state of the virusinvaded host cells. In this method, anti-RSV-antibody modified gold nanoparticles(Au NPs) could bind with the invading virus on the cell membrane of the infected host cells through the specific antibody-antigen binding. Then,the host cells could be imaged by the localized surface plasmon resonance light scattering properties of Au NPs under a dark-field light scattering microscopy, which could be further used to semi-quantify the invading virus.展开更多
基金This work was supported by a grant from the Natural Science Foundation of Nenan Province, China （No.102300410078）.
文摘OBJECTIVE To investigate the available parameters in gynecological screening for cervical lesions by liquid-based cytology technology （ThinPrep Cytology Test, TCT） and The Bethesda System （TBS）, also with computer image analysis. METHODS With application of the image analysis system, all grades of cervical lesion cells were detected quantitatively and sorted in atypical squamous cells of undetermined significance （ASCUS）, atypical squamous cells-cannot exclude HSIL （ASC-H）, low-grade squamous intraepithelial lesion （LSIL）, high-grade squamous intraepithelial lesion （HSIL） and cervical squamous cell carcinoma （SCC） with the mean optical density （MOD）, average grey （AG）, positive units （PU）, and nucleus to cytoplasmic ratio （N： C）. Differences between each group of cells were compared and analyzed statistically. RESULTS Apart from four stereologic parameters in LSIL and HSIL groups there were no differences among them, in the other groups, there was statistically significant in differences between MOD, AG and PU values. Differences between them in the ratio of nucleus to cytoplasm were highly statistically significant. CONCLUSION Stereological indexes may serve as a screening tool for cervical lesions. The image analysis system is expected to become a new means of cytological assisted diagnosis.
基金The General Foundation of Tianjin Science Committee for Applied Basic Research (08JCZDJC21000)Chinese Ministry of Education (30770081)
文摘In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.
文摘The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with which the Th cells status of the patients with gynaecological tumors and chronic renal failure （CRF） can be analyzed. IL-2 eDNA and IL-4 eDNA were prepared, and the plasmid pMD18 carrying IL-2 eDNA or IL-4 eDNA fragment was constructed and cloned as the template for quantitative determination. The primers and probes labelled with 6-carbexy-fluorescein （FAM） and 6-carboxy- tetramethylrhodamine （TAMRA） were prepared, and the experimental conditions were optimized to set up the FQ-RT-PCR method for quantitative determination of IL-2 mRNA and IL-4 mRNA. Th cells enriched from peripheral blood mononuclear cells （PBMCs） of 20 healthy volunteers （HVs）, 16 gynaecological benign （GB） cases, 18 gynaecological malignant （GM） tumor cases and 16 chronic renal failure （CRF） patients were tested for IL-2 mRNA and IL-4 mRNA by FQ-RT-PCR. The house-keeping gene β-actin was used as the internal control gene of the experiment. The standard curve for log concentration of series of quantitative templates vs threshold cycle （CT） was established by linear regression, and the linear range was 102-107 copies/μl. The imprecision test showed the CV of inter-assay and intra-assay of a high cont- ent sample by FQ-RT-PCR were 7.8% and 12.5%, respectively. The CV of inter-assay and intra-assay of a low content sample were 10.8% and 19.5%, respectively. The IL-2 mRNA expressions in Th of the patients with gynaecological malignant tumor （compared with the HVs and the patients with gynaecological benign disease） and in Th of the CRF patients （compared with the HVs） were declined significantly and at the same time the IL-4 mRNA expression increased significantly （ P 〈 0. 001 ）. A simple, sensitive and accurate FQ-RT-PCR method for the quantitative detection of IL-2 mRNA and IL-4 mRNA has been established. The IL-2 mRNA and IL-4 mRNA expressions in Th cells of the patients with gynaecological malignant tumor and the CRF patients were polarized and displayed Th2 response. It suggests that the function of the Th cells of the patients with gynaecological malignant tumor or CRF is at unbalance.
基金supported by the National Basic Research Program of China(2011CB933600)Chongqing Fundamental and Advanced Research Project(cstc2013jcyj A50008)the Fundamental Research Funds for the Central Universities(XDJK2015B029)
文摘A novel sensitive semi-quantitative virus detection technique was developed using the respiratory syncytial virus(RSV) as an example, through dark-field light scattering imaging of the surface state of the virusinvaded host cells. In this method, anti-RSV-antibody modified gold nanoparticles(Au NPs) could bind with the invading virus on the cell membrane of the infected host cells through the specific antibody-antigen binding. Then,the host cells could be imaged by the localized surface plasmon resonance light scattering properties of Au NPs under a dark-field light scattering microscopy, which could be further used to semi-quantify the invading virus.
