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1.
Article in Chinese | WPRIM | ID: wpr-695634

ABSTRACT

Objective·To explore the effects of adenovirus vector-mediated small interfering RNA (siRNA) targeting phosphatase nuclear targeting subunit (PNUTS) on proliferation,invasion,migration and epithelial-mesenchymal transition (EMT) of laryngeal squamous cell carcinomas Hep-2 cells and its mechanism.Methods· Recombinant adenovirus vector expressing PNUTS siRNA was infected into laryngeal squamous cell carcinomas Hep-2 cells and the experiment was designed into PBS group,Ad-GFP group and Ad-siPNUTS group.Levels ofPNUTS mRNA and protein were detected by real-time PCR and Western blotting respectively.MTT assay was used to detect proliferation abilities of Hep-2 cells.Transwell assays were used to detect invasion and migration abilities of Hep-2 cells.The expression levels of total Rb,phosphorylated Rb (p-Rb),PI3K,phosphorylated AKT (p-AKT),E2FI,E-cadherin,N-cadherin and ZEB1 protein were detected by Western blotting.Results· Compared with Ad-GFP group,in Ad-siPNUTS group,the PNUTS mRNA and protein (both P=0.000) levels were dramatically decreased.The proliferation of Ad-siPNUTS infected Hep-2 cells were inhibited on the second day (P=0.004),the third day (P=0.001) and the fourth day (P=0.000).Meanwhile,the invasion and migration abilities of Ad-siPNUTS infected Hep-2 cells were decreased (both P=0.000).The expression levels of total Rb (P=0.000),p-Rb (P=0.000),PI3K (P=0.023),p-AKT (P=0.000),E2F 1 (P=0.000),N-cadherin (P=0.005) and ZEB 1 (P=0.000) were decreased while the E-cadherin (P=0.003) was increased.Conclusion· Ad-siPNUTS could inhibit the proliferation,invasion and migration abilities of Hep-2 cells and reverse the development of EMT,which may be related to PI3K/AKT signaling pathway and Rb signaling pathway.

2.
Journal of Experimental Hematology ; (6): 1093-1100, 2018.
Article in Chinese | WPRIM | ID: wpr-689523

ABSTRACT

<p><b>OBJECTIVE</b>To detect whether the murine T-cell lymphoma cell line EL4 could be infected by murine cytomegalovirus (MCMV), and to observe the morphological changes and apoptosis of El4 cells before and after infection.</p><p><b>METHODS</b>EL4 cells were infected with MCMV smith strain with 1, 10 and 100 multiplicity of infection (MOI) respectively. The morphology of the cells was observed by light microscopy and Wright's-Giemsa staining. The survival rate was calculated by trypan blue staining. RT-PCR-based assay was used to detect the copy number of MCMV-DNA in the infected ELA cells. Flow cytometry was used to detect apoptosis. RT-qPCR was used to detect the mRNA expression levels of P53, P21, cFlip and Caspase 8. The protein expression levels of Caspase8, P53, BAX, BCL-2 and Cleaved Caspase3 proteins were detected by Western blot.</p><p><b>RESULTS</b>After Wright-Giemsa staining, it was found that the infected EL4 cells displayed larger volume, irregular nuclei and the folded twist under light microscopy. Compared with the normal control group, the survival rate of EL4 cells decreased, and the apoptosis rate statistically significantly (P<0.05) increased with the increasing MOI and the infected time in each group. While, the level of apoptosis protein P53, BAX/BCL-2, Cleaved-caspase3 and Caspase8 were up-regulated. And the survival rate, apoptosis rate and the apoptosis protein level of infected EL4 cells with MOI=10 were the most obvious at the 5day. Compared with MCMV infection group (MOI=60), the content of MCMV DNA in EL4 cells was decreased in MCMV+GGV group [MOI=60, GCV 25 (g/ml)], and the cell apoptosis rate and apoptosis protein expression of P53, Caspase8, BAX/BCL-2 were decreased (P<0.05).</p><p><b>CONCLUSION</b>Murine T-cell lymphoma cell line EL4 can be infected by MCMV and displayes an obvious apoptosis phenomenon. MCMV may up-regulate the expression levels of apoptosis protein P53, BAX-BCL-2, Cleaved-caspase3 and Caspase8 in EL4 cells. The drug ganciclovir reduces the copy mumber of MCMV DNA in infected EL4 cells and inhibited the killing effect of MCMV on EL4 cells.</p>


Subject(s)
Animals , Apoptosis , Caspase 8 , Ganciclovir , Lymphoma, T-Cell , Mice , Muromegalovirus
3.
Article in English | WPRIM | ID: wpr-812511

ABSTRACT

The present study investigated the chemical constituents of the roots of Stellera chamaejasme (Thymelaeaceae). One new biflavone glucoside (1), along with other thirteen known compounds (2-14), was isolated by repeated column chromatographic methods and their structures were elucidated on the basis of spectral analyses. The cytotoxic activities of selected compounds were evaluated against four human cancer cell lines (A549, BEL-7402, HCT-116, and MDA-MB-231) by the SRB assay method. Compound 9 showed remarkable cytotoxicity against BEL-7402 with IC50 value being 0.65 μg·mL(-1); compounds 7, 8, and 12 exhibited significant cytotoxic activity against A549 with IC50 values being 2.38, 1.57, and 2.35 μg·mL(-1), respectively.


Subject(s)
Antineoplastic Agents, Phytogenic , Chemistry , Pharmacology , Therapeutic Uses , Biflavonoids , Chemistry , Pharmacology , Cell Line, Tumor , Glucosides , Chemistry , Pharmacology , Humans , Inhibitory Concentration 50 , Molecular Structure , Phytotherapy , Plant Extracts , Chemistry , Pharmacology , Therapeutic Uses , Plant Roots , Chemistry , Thymelaeaceae , Chemistry
4.
Chinese Journal of Hematology ; (12): 605-609, 2012.
Article in Chinese | WPRIM | ID: wpr-278359

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the regular pattern of Cytomegalovirus (CMV)-specific T cells (CTL) immune reconstitution after human leukocyte antigen (HLA) matched sibling donor allogeneic bone marrow(BM) plus peripheral blood hematopoietic stem cell (PBSC) transplantation.</p><p><b>METHODS</b>CTL from seventeen patients after transplantation was detected by flow cytometry, the IFN-γ secretion ability of CTL by enzyme-linked immunospot (ELISPOT) assay, and clonal analysis of TCR Vβ subfamily by gene scan assays. The relationship between CTL reconstitution and CMV infection was studied.</p><p><b>RESULTS</b>Both number and function of recipients CTL reached to normal control level at 30 d post-transplantation. The recipients achieved a high frequency CTL with IFN-γ response and restoration of T-cell receptor β (TCR Vβ) repertoire at one year post-transplantation. CTL with the central memory CD45RO(+)CD62L(+) cell phenotype expanded in PB when CMV was reactivated. The incidence of CMV reactivation was 35.83% (17.91% - 63.10%) after transplantation, and none of them developed CMV disease.</p><p><b>CONCLUSION</b>After HLA matched related donor transplantation using mixed grafts, immune recovery to CMV seems to be early and fast. The incidence of CMV infection and disease are low.</p>


Subject(s)
Adult , Cytomegalovirus , Allergy and Immunology , Female , HLA Antigens , Allergy and Immunology , Hematologic Diseases , Allergy and Immunology , General Surgery , Hematopoietic Stem Cell Transplantation , Humans , Male , Middle Aged , Peripheral Blood Stem Cell Transplantation , Siblings , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Tissue Donors , Young Adult
5.
Chinese Journal of Epidemiology ; (12): 413-417, 2012.
Article in Chinese | WPRIM | ID: wpr-269144

ABSTRACT

Objective To investigate the prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong province.Methods Data from 169 218pregnant women in different regions of Guangdong province from January 2005 to June 2010 were analyzed retrospectively.The prevalence and epidemiological characteristics of thromboembolic disease during pregnancy or puerperium were investigated.Results Of the studied population,( 1 )20 l cases ( 1.3‰ ) suffered from thromboembolic disease during pregnancy or puerperium including 128 cases of deep vein thrombosis (DVT),68 cases of cerebral venous thrombosis (CVT) and 5pulmonary embolism,the prevalence rates were 0.8‰,0.4‰,and 0.02‰ respectively.(2) Risk factors in different regions showed that,in the Pearl River Delta area,the major risk factors for DVT would include previous or family history of thrombosis,pregnancy complications,with medically involved diseases,prolonged bed rest and pregnancy weight gain > 15 kg etc.While in castern,western,northern parts of Guangdong,the major risk factors for DVT would include pregnancy weight gain > 15 kg,prolonged bed rest,preeclampsia,cesarean section and complications during pregnancy.In Pearl River Delta region,the major risk factors for CVT would include eclampsia,preeclampsia,pregnancy complications,prolonged bed rest >3 days,past history or family history of thrombosis.While eclampsia,preeclampsia,advanced age or younger age,pregnancy weight gain >15 kg,complications during pregnancy were the major risk factors for CVT in the eastern,western or northem parts of Guangdong.Conclusion Prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong were different.It was crucial to take effective measures in pregnant women with different epidemiological characteristics and risk factors to prevent and reduce the incidence of peripartum thromboembolic disease.

6.
Journal of Experimental Hematology ; (6): 1316-1320, 2009.
Article in Chinese | WPRIM | ID: wpr-343295

ABSTRACT

The aim of study was to investigate the modulation effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on adhesion molecule expression of memory T lymphocyte in the bone marrow grafts. rhG-CSF was administered in 41 donors by subcutaneous injection for 5 consecutive days. Bone marrow grafts were collected on day 4. The percentages of CD4+ and CD8+, and the expressions of CD49d, CD54, CD62L and CD11a on donor T cells of steady state-bone marrow grafts (SS-BM, n=11) and rhG-CSF primed bone marrow (G-BM, n=30) were analyzed by using multi-color flow cytometry. The results indicated that the percentages of CD4+ and CD8+ T cells were significantly lower in G-BM than those in SS-BM (p<0.05). There were no significant differences in the percentages of memory CD4+ and CD8+ T cells between SS-BM and G-BM (p>0.05). The expressions of CD49d on CD4+ and CD8+T cells were significantly lower in G-BM than that in SS-BM (p<0.05). Compared with SS-BM, the expressions of CD54 on CD4+, memory CD4+ T cells and CD8+ T cells were significantly lower in G-BM (p<0.05). The expressions of CD62L on CD4+ and CD8+ T cells and memory T cells were all significantly lower in G-BM (p values were all less than 0.001). The expressions of CD11a on CD4+, memory CD4+ T cells were significantly lower in G-BM than that in SS-BM (p<0.05). There were no significant differences in the expression of CD11a on CD8+, memory CD8+ T cells between SS-BM and G-BM (p>0.05). It is concluded that the expression of cell adhesion molecules on the CD4+ and CD8+ T cells in G-BM is down-regulated after rhG-CSF treatment of healthy donors.


Subject(s)
Adolescent , Adult , Aged , Bone Marrow , Bone Marrow Cells , Allergy and Immunology , Metabolism , Bone Marrow Transplantation , Allergy and Immunology , Methods , Cell Adhesion Molecules , Metabolism , Female , Granulocyte Colony-Stimulating Factor , Pharmacology , Humans , Living Donors , Male , Middle Aged , Recombinant Proteins , T-Lymphocytes , Allergy and Immunology , Metabolism , Young Adult
7.
Article in Chinese | WPRIM | ID: wpr-267927

ABSTRACT

The study was aimed to analyze the difference of naive and memory CD4(+) and CD8(+) T-cell subsets between steady-state bone marrow (SS-BM) and recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood grafts (G-PB). Four CD4(+) and CD8(+) T-cell subsets classified according to the expression of CD45RA and CD62L were determined by three-color flow cytometry. The results showed that the percentage of CD4(+), CD8(+) T-cell subsets and the ratios of CD4/CD8 in G-PB were significantly higher than those in SS-BM (p < 0.05). The percentage of CD4(+) naive T-cells in G-PB was significantly lower than that in SS-BM (p < 0.001). As compared with SS-BM, the percentage of CD4(+) effector memory T-cells was significantly high in G-PB (p < 0.001). There were no significant differences in the percentages of the four CD8(+) T-cell subsets between SS-BM and G-PB (p > 0.05). The percentage of CD4(+)CD62L(+) T-cells in G-PB was significantly lower than that in SS-BM (p = 0.001). The absolute numbers of CD4(+) and CD8(+) T-cell subsets, the eight naive and memory CD4(+) and CD8(+) T-cell subsets were significantly higher in G-PB than those in SS-BM (p < 0.001). It is concluded that the difference of naive and memory CD4(+) and CD8(+) subsets between G-PB and SS-BM may partially explain why the incidence and severity of acute graft-versus-host disease (GVHD) was similar and the incidence of chronic GVHD was different after transplantation with SS-BM or G-PB.


Subject(s)
Adolescent , Adult , Aged , Bone Marrow Transplantation , CD4-CD8 Ratio , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Female , Graft vs Host Disease , Granulocyte Colony-Stimulating Factor , Therapeutic Uses , Hematopoietic Stem Cell Mobilization , Humans , Male , Middle Aged , Peripheral Blood Stem Cell Transplantation , Recombinant Proteins , Young Adult
8.
Article in Chinese | WPRIM | ID: wpr-267926

ABSTRACT

The study was purposed to investigate the effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in healthy donors after recombinant human granulocyte colony-stimulating factor (rhG-CSF) application in vivo. Sixty-two healthy donors were treated with rhG-CSF [5 microg/(kg.d)] injected subcutaneously for five consecutive days. Donors were divided into groups A and B, and 31 donors were there in each group. Bone marrow grafts and peripheral blood stem cell grafts were harvested on the day 4 and 5 respectively in group A. In group B, only peripheral blood grafts were collected on both the day 4 and 5. The quantities of the cell components, CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry. The results showed that median counts of nuclear cells (1.56 x 10(5)), lymphocytes (8.56 x 10(4)), CD3(+) (6.12 x 10(4)), CD3(+)CD4(+) (3.38 x 10(4)), CD3(+)CD8(+) (2.27 x 10(4)), CD14(+) (3.83 x 10(4)), CD34(+) cells (744) and CD3(+)CD4(-)CD8(-) T cells (3588) per microliter of peripheral blood stem cell grafts in group A were similar to counts of nuclear cells (1.40 x 10(4)), lymphocytes (7.34 x 10(4)), CD3(+) (5.32 x 10(4)), CD3(+)CD4(+) (3.06 x 10(+)), CD3(+)CD8(+) (1.83 x 10(4)), CD14(+) (3.21 x 10(4)), CD34(+) cells (554) and CD3(+)CD4(-)CD8(-) T cells (3120) in group B (p > 0.05). There were no difference in the ratios of CD4(+) cells/CD8(+) cells, CD14(+) cells/CD3(+) cells and CD3(+)CD4(-)CD8(-) T cells/CD3(+) cells in peripheral blood stem cell grafts between group A [1.52 (0.54 - 2.87)], [0.57 (0.15 - 1.64)], [0.064 (0.018 - 0.673)] and group B [1.68 (0.31 - 3.35)], [0.59 (0.18 - 1.25)], [0.063 (0.021 - 0.136)] (p > 0.05). It is concluded that no effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in the same donor is found after rhG-CSF application in vivo, bone marrow grafts and peripheral blood stem cell grafts can be collected respectively or simultaneously.


Subject(s)
Adolescent , Adult , Bone Marrow Cells , Cell Biology , Bone Marrow Transplantation , Female , Granulocyte Colony-Stimulating Factor , Therapeutic Uses , Hematopoietic Stem Cell Mobilization , Humans , Male , Middle Aged , Peripheral Blood Stem Cell Transplantation , Recombinant Proteins , Tissue Donors , Young Adult
9.
Chinese Journal of Hematology ; (12): 512-516, 2008.
Article in Chinese | WPRIM | ID: wpr-239990

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the cell composition in granulocyte colony-stimulating factor One (rhG-CSF) primed bone marrow grafts (G-BM) from donors with different characteristics.</p><p><b>METHODS</b>hundred and fifty healthy donors were injected subcutaneously rhG-CSF (5 microg x kg(-1) x d(-1)) in five consecutive days. Bone marrow and peripheral blood stem cells were harvested at day 4 and day 5. The number of CD3, CD3+ CD4+, CD3+ CD8+, CD14+ , CD34+ and CD3+ CD4- CD8- cells were determined by multicolor flow cytometry. Cell composition of G-BM from donors with different characteristics, including sex, age, weight, pregnancy were analysed.</p><p><b>RESULTS</b>The absolute number of nuclear cells (NCs), lymphocytes, CD3+, CD3+ CD4+, CD3+ CD8+, CD14+, CD34+ and CD3+ CD4- CD8- cells in G-BM were 31 050 (12 200 -58 100), 2122 (506 - 6618), 1344 (307 - 4791), 692 (145 - 3038), 616 (112 - 2575), 986 (265 -2958), 63 (11 -505) and 83 (9 -390) per microliter, respectively. The number of NCs [33 800 (18 600 - 57 100)], CD34+ cells [76 (22 -505)], and CD3+ CD4+ CD8- regulatory T cells [97 (11 - 380)] harvested from younger donors (aged < or = 38 years) were significantly higher than those from older ones (aged > 38) [28000 (12200-58100), 49 (11-220), and 65 (9 - 389) per microliter (P = 0.027, < 0.001 and = 0.001, respectively)]. Compared with that in donors with peripheral blood monocytes (PBMs) < or = 0.37 x 10(9)/L, higher number of NCs in G-BM [27 150 (13 800 - 58 100) vs 33 550 (12 200 - 57 100), P = 0.005] were collected in donors with PBMs > 0.37 x 10(9)/L. Multivariate analysis showed that donors age (< or = 38 vs > 38 years; P = 0.01) and monocyte number in peripheral blood (< or = 0.37 x 10(9)/L vs > 0.37 x 10(9)/L; P = 0. 003) were factors predicting for NC yields, and donors age ( < or = 38 vs > 38 years; P < 0.001) was factor predicting for yields of CD34+ cells (P < 0.001) and CD3+ CD4- CD8- regulatory T cells (P = 0.001) collection.</p><p><b>CONCLUSION</b>Donor age is a factor for the yields of NCs, CD34+ cells, and CD3+ CD4- CD8- regulatory T cells collection in G-BM, and donor's PBMs more than 0.37 x 10(9)/L, is another factor affecting NCs harvests.</p>


Subject(s)
Adolescent , Adult , Age Factors , Antigens, CD34 , Bone Marrow Cells , Allergy and Immunology , Female , Granulocyte Colony-Stimulating Factor , Therapeutic Uses , Hematopoietic Stem Cell Mobilization , Humans , Male , Middle Aged , Recombinant Proteins , T-Lymphocyte Subsets , Allergy and Immunology , Tissue Donors , Young Adult
10.
Article in Chinese | WPRIM | ID: wpr-282771

ABSTRACT

To investigate expression of plk-1 gene and PLK-1 protein in acute leukemia patients and its clinical significance, and to observe the distribution of PLk-1 protein in acute leukemin cells, the mononuclear cells were separated from the bone marrow or peripheral blood of acute leukemia patients, bone marrow benign proliferation individuals and normal individuals. The expression of plk-1 gene and PLK-1 protein in those cells were detected with RT-PCR and flow cytometry respectively, the distribution pattern of PLK-1 was observed by fluorescent inverted microscope. The result showed that the expressions of plk-1 gene and PLK-1 protein in mononuclear cells of acute leukemia patients were much higher than that of bone marrow benign proliferation individuals and normal individuals. Fluorescent inverted microscopy revealed that PLK-1 was highly concentrated in cytoplasm of acute leukemia cells during interphase of mitosis, and it was found that PLK-1 was mainly distributed between sister chromatid during the mitosis in mononuclear cells of acute leukemia patients, but the expressions of plk-1 gene and PLK-1 protein almost were not observed in cells of benign proliferative bone marrow and normal bone marrow. It is concluded that increased plk-1 gene and protein perhaps play an important role in abnormal proliferation of acute leukemia cells and correlate with the malignamcy of leukemia. plk-1 gene or PLK-1 protein may be considered as a new target of therapy, and one of useful indicators in evaluation of curative efficiency and prognosis.


Subject(s)
Adolescent , Adult , Aged , Biomarkers, Tumor , Genetics , Bone Marrow Cells , Metabolism , Cell Cycle Proteins , Genetics , Child , Female , Humans , Leukemia, Myeloid, Acute , Genetics , Metabolism , Male , Middle Aged , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Metabolism , Prognosis , Protein-Serine-Threonine Kinases , Genetics , Proto-Oncogene Proteins , Genetics
11.
Chinese Journal of Hepatology ; (12): 190-193, 2005.
Article in Chinese | WPRIM | ID: wpr-349172

ABSTRACT

<p><b>OBJECTIVES</b>To study the relationship between CT imaging features and the severity of chronic viral hepatitis.</p><p><b>METHODS</b>The clinical data and laboratory information of 120 chronic virus hepatitis patients were reviewed retrospectively. They were categorized into mild (34 cases), moderate (22), severe hepatitis (26) and hepatitis gravis (38) groups based on international standardized clinical criteria. All patients underwent standardized contrast-enhanced spiral CT dual-phase scanning of the upper abdomen. The changes of the liver, bile duct, spleen, portal venous system, lymph nodes of the upper abdomen, peritoneal cavity and pleural cavity were examined and noted. Correlation analysis was applied for the relationship of CT imaging features and the severity indices of chronic virus hepatitis.</p><p><b>RESULTS</b>As the severity of chronic hepatitis increased, which was reflected as serum albumin level went down and the value of ALT went up, the incidences of the following manifestations were also intensified: (1) intrahepatic perivascular lucency, (2) edema of the gallbladder fossa, (3) enlargement of abdominal lymph nodes, (4) ascites and (5) thoracic changes. Correlation between these CT manifestations and clinical severity of chronic viral hepatitis was of statistical significance.</p><p><b>CONCLUSION</b>A close correlation exists between certain CT features of chronic hepatitis and the clinical severity of chronic viral hepatitis.</p>


Subject(s)
Adolescent , Adult , Aged , Female , Hepatitis B, Chronic , Diagnostic Imaging , Humans , Liver , Diagnostic Imaging , Male , Middle Aged , Retrospective Studies , Severity of Illness Index , Tomography, X-Ray Computed
12.
Article in Chinese | WPRIM | ID: wpr-679734

ABSTRACT

Objective To investigate the clinical and mammographic features of plasma cell mastitis.Methods Twenty-five patients(28 lesions)with histologically confirmed plasma cell mastitis, aged from 26 to 70 years(mean age 41 years),were examined with X-ray mammography.The clinical manifestations and imaging features were retrospectively reviewed.Results No case was in lactation.The painful irregular masses,ranged from 1.3 to 8cm in size,were found in 22 patients,while 3 patients with acute episode.Recurrent episodes of breast masses were noted in 4 patients.Based on the mammographic appearances,the plasma cell mastitis were classified as the following four types:inflammation-like type (2/28),ductal ectasia type(3/28),focal infiltration type(10/28)and nodular type(13/28).The valuable radiogyaphic signs:(1)An asymmetrically increased density along the lactiferous duct with a flame-like appearance,inhomogeneous low density tubular structures and scattered stick-shape calcifications.(2) Architectural distortion and oil cysts formation in adjacent area,(3)Subareolar ductal ectasia.Conclusions The clinical and mammographic characteristics of plasma cell mastitis are critical to avoiding unnecessary surgery.Histopathological result is needed for the diagnosis in patients highly suspected of malignancy.

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