Coconut(Cocos nucifera L.),a major oil and fruit crop of the Arecaceae family,is extensively cultivated across the Asia—Pacific region.Despite its agricultural importance,genome assembly in coconut remains challengin...Coconut(Cocos nucifera L.),a major oil and fruit crop of the Arecaceae family,is extensively cultivated across the Asia—Pacific region.Despite its agricultural importance,genome assembly in coconut remains challenging due to its large genome size and high proportion of repetitive sequences.Allele-specific expression(ASE)plays a key role in regulating plant development and evolution,yet research on ASE in coconut is limited(Shao et al.,2019;Li et al.,2021;Zhang et al.,2021;Hu et al.,2022).Among phenotypic traits,fruit color is especially important as an indicator of maturity,guiding harvest timing and post-harvest processes(Kapoor et al.,2022).While prior studies have explored various coconut traits such as salt tolerance,fiber content,and plant height(Wang et al.,2021;Yang et al.,2021),investigations into ASE and fruit color remain scarce.展开更多
In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl 2) in peripheral blood lymphocytes (PBLC) and the level of IFN γ and IL 4 in serum of the patients with...In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl 2) in peripheral blood lymphocytes (PBLC) and the level of IFN γ and IL 4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl 2 in PBLC and the level of IFN γ and IL 4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl 2 was significantly lower (both P< 0.01). The level of IFN γ in serum of CA was significantly lower than in the normal control group ( P< 0.01), but IL 4 was significantly lower (both P< 0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN γ in serum of patients with CA, but had a positive correlation with the level of IL 4; The expression of Bcl 2 had a positive correlation with the level of IFN γ, but had a negative correlation with the level of IL 4. All the correlation coefficients had significant differerce by t test ( P< 0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the THl associated cytokines (eg: IFN γ) and the increased secretion of the TH2 asssociated cytokines (eg: IL 4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.展开更多
Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the ty...Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the types of IFN, which suppresses proliferation and induces apoptosis in some cell types while inhibiting apoptosis in others. The aim of this study was to explore the effect of IFNα-2a on Fas expression and the apoptosis rate of peripheral blood cytotoxic T cells (CTLs) in patients with hepatitis B. METHODS:Peripheral blood mononuclear cells were isolated from 26 patients with hepatitis B including 16 patients with chronic hepatitis B and 10 patients with chronic severe hepatitis B. Fas expression and apoptosis rate of CTLs were analyzed with flow cytometry before and after IFNα-2a treatment. RESULTS:Before IFNα-2a treatment, Fas expression and apoptosis rate of CTLs from patients with chronic hepatitis B were significantly higher than those from patients with chronic severe hepatitis B and healthy controls respectively. No significant difference was observed between Fas expression and apoptosis rate of CTLs from patients with chronic severe hepatitis B and healthy controls. After IFNα-2a treatment,Fas expression and apoptosis rate of CTLs from different groups were compared with those before IFNα-2a treatment, showing no significant difference despite alternation of different degree. CONCLUSIONS:Activation induced cell death (AICD) exists in peripheral blood CTLs from patients with hepatitis B. No effect of IFNα-2a exerts on Fas expression and apoptosis rate of Fas in patients with hepatitis B.展开更多
Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon ...Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon cancer cell line CCL229. Methods: Using the molecular and biochemical techniques and electron microscopy to analyze the expression of iNOS, production of NO and growth characteristics of human colon cancer cells. Results: cytokine treatment can induce expression of the iNOS gene and production of nitric oxide was significantly higher after treatment of CCL229 cells with TNF-αor IFN-γ. Treatment with either cytokine or a combination of both significantly increased levels of Malondialdehyde (MDA) over control. Furthermore, cytokine treatment increased the proliferation inhibition rate as assessed in vitro and decreased the cell proliferation index on flow cytometry. Electron microscopy showed that cells treated with cytokines had fewer pseudopodia or cell processes than control cells and that cytokine treated cells had dilatation of the mitochondria and endoplasmic reticulum and dilated vesicular or tubular cisternae. Conclusion: Our findings indicate that TNF-α and IFN-γ induce the expression of iNOS gene in CCL229 cells, which increases the production of nitric oxide, inhibits proliferation, causes lipid peroxidation, and results in ultrastructural changes.展开更多
Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and c...Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and cultured in vitro, and the expression of IL-10R1 and IL-10R2 gene was analyzed after cells had been treated with TH2-type cytokines IL-10 and TH1-type cytokines IFN-γ within 60 rain with semiquantitative reverse transcriptase-PCR, then the influence of IL-10 and IFN-γ on expression of IL-10R protein was examined by first trimester DSC using flow cytometry. In addition, the vitality of DSC was detected by MTT. Results IL-10R1 mRNA levels of DSC treated with IL-10 （10 ng/ml） reached the peak level within 15 rain, and were significantly lower at 30 rain, then were not detected at 45 min. The expression of IL-10R1 were induced to moderate level by IFN-γ（10 ng/ml） within 30 rain, and reduced to undetected levels at 60 min. There was no significant difference of IL-10R2 expression （P〉0.05） between treated and not with the abovementioned cytokines. The IL-10R protein expression and vitality of DSC were significantly enhanced by IL-10 （10 ng/ml） and IFN-γ （10 ng/ml） which treated DSC 48 h （P〈0.05）. Coneclusion IL-10 and IFN-γ may play an important role of biologic function in early pregnancy by influencing IL-10R expression of DSC.展开更多
Because yeast codon preferred to synthesize the gene sequence of ChIFN-α mature peptide, Pichia pastoris expression system was connected into vector pPICgK, to construct recombinant plasmid pPIC9K-α. After digestion...Because yeast codon preferred to synthesize the gene sequence of ChIFN-α mature peptide, Pichia pastoris expression system was connected into vector pPICgK, to construct recombinant plasmid pPIC9K-α. After digestion and linearization, the recombinant vector was cloned into P. pastor/s GS115, high copy transformant and its methanol utilization type were screened, and its expression conditions were optimized. The results showed that ChlFN-α successfully expressed in P. pastor/s, the expression amount of recombinant protein was the largest after induced expression in 0.5 % methanol containing 2% acid hydrolyzed casein at 28 ℃ for 72 h, and its antiviral activity was the highest.展开更多
In ord,r to study the relatiouship between the expression or PML and the antiproliferative effect or interferon (IFN), prostate cancer cells, PC-3, DU145 and LNCaP were cultured, andtreated with IFN-α. The cell growt...In ord,r to study the relatiouship between the expression or PML and the antiproliferative effect or interferon (IFN), prostate cancer cells, PC-3, DU145 and LNCaP were cultured, andtreated with IFN-α. The cell growth. rates were measured by crystal violet assay, and the expressionof PML was detected by immunofluomcent staining with PML Polyclonal antibody. Our study indicated that the growth rates or PC-3 and DU145 were significantly inhibited by IFN-α, whereas nosuch effect can be found on LNCaP cell line. The immunofluorescent staining showed that both thestaining intensity and the number or the PML nuclear boles were significantly increased in PC-3 andDU145 cells, however, no significant changes in the PML nuclear body were found in the IFs-αtreated LNCaP cells. Our study demeastrated that the antiproliferative,effct of IFN in prostate cancer cells may be directly related to the induced expression of PML in response to its treatment. Ourfinding supports the hypothesis that PML may mediate some antiproliferative erfects of IFN.展开更多
Objective： This study investigated the effects of rh-TRAIL with or without chemotherapeutic drugs on the apoptosis of the osteosarcoma cell line, MG-63, and the influence of chemotherapeutic drugs on changes in the e...Objective： This study investigated the effects of rh-TRAIL with or without chemotherapeutic drugs on the apoptosis of the osteosarcoma cell line, MG-63, and the influence of chemotherapeutic drugs on changes in the expression of DR5 and YinYang 1 （YY1） in MG- 63 cells. Methods： The effects of treatment with rh-TRAIL alone and/or chemotherapetic drugs on MG-63 cell growth inhibition and apoptosis were measured using the MTT assay, FACS analysis ofAunexin V labeled cells, and the mRNA changes of DR5 and YY1 were detected by RT-PCR. Results： Rh-TRAIL protein inhibited the growth of MG-63 cells, and this inhibition was increased by adriamycin and IFN-γ. Adriamycin and IFN-γ significantly facilitated the induction of the expression of DR5 and reduced the expression of YY1. Conclusion： The apoptosis-inducing effect of rh-TRAIL in MG-63 cells was enhanced by chemotherapeutic drugs.展开更多
AIM： To study the association between host immunity and hepatitis B virus （HBV） recurrence after liver transplantation. METHODS： Peripheral blood mononuclear cells （PBMC） were isolated from 40 patients with hepa...AIM： To study the association between host immunity and hepatitis B virus （HBV） recurrence after liver transplantation. METHODS： Peripheral blood mononuclear cells （PBMC） were isolated from 40 patients with hepatitis B and underwent orthotopic liver transplantation （OLT） before and 2, 4, 8 wk after surgery. After being cultured in vitro for 72 h, the levels of INF-γ and TNF-α in culture supematants were detected with ELISA. At the same time, the quantities of HBV DNA in serum and PBMCs were measured by real time PCR. RESULTS： The levels of INF-γ and TNF-α in PBMC culture supernatants decreased before and 2, 4 wk after surgery in turns （INF-γ 155.52±72.32 ng/L vs 14.76±9.88 ng/L vs 13.22±10.35 ng/L, F= 6.946, P= 0.027〈0.05; TNF-α 80.839±46.75 ng/L vs18.59±17.29 ng/L vs9.758±7.96 ng/L, F= 22.61, P = 0.0001〈0.05）. The levels of INF-γ and TNF-α were higher in groups with phytohemagglutinin （PHA） than in those without PHA before surgery. However, the difference disappeared following OLT. Furthermore, INF-γ and TNF-α could not be detected in most patients at wk 4 and none at wk 8 after OLT. The HBV detection rate and virus load in PBMC before and 2, 4 wk after surgery were fluctuated （HBV detected rate： 51.4%, 13.3%, 50% respectively; HBV DNA： 3.55±0.674 log（10） copies/mL vs 3.00±0.329 log（10） copies/mL vs 4.608±1.344 log（10） copies/mL, F= 7.582, P= 0.002〈0.05）. HBV DNA in serum was 4.48±1.463 log（10） copies/mL before surgery and 〈 10^3 copies/mL after OLT except for one with 5.72×10^6 copies/mL 4 wk after OLT who was diagnosed as HBV recurrence. The levels of INF-γ and TNF-α were lower in patients with a high HBV load than in those with a low HBV load （HBV DNA detected/undetected in PBMCs： IFN-γ 138.08±72.44 ng/L vs 164.24±72.07 ng/L, t = 1.065, P= 0.297〉0.05, TNF-α 80.75±47.30 ng/L vs 74.10±49.70 ng/L, t= 0.407, P= 0.686〉 0.05; HBV DNA positive/negative： IFN-γ 136.77±70.04 ng/L vs 175.27±71.50 ng/L, t= 1.702, P= 0.097〉0.05; TNF-α 75.37±43.02 ng/L vs 81.53±52.46 ng/L, t = 0.402, P = 0.690〉0.05）. CONCLUSION： The yielding of INF-γ and TNF-α from PBMCs is inhibited significantly by immunosuppressive agents following OLT with HBV load increased, indicating that the impaired immunity of host is associated with HBV recurrence after OLT.展开更多
Objective:To investigate the effects of interferon-a (IFN-a) and IFN-a combined with interleukin-6 (IL-6) on growth and expression of bcr-abl, bcl-2 and c--myc genes in the mononuclear cells (MNCs) from bone marrow (B...Objective:To investigate the effects of interferon-a (IFN-a) and IFN-a combined with interleukin-6 (IL-6) on growth and expression of bcr-abl, bcl-2 and c--myc genes in the mononuclear cells (MNCs) from bone marrow (BM) of patients with chronic myelogenous leukemia (CML). Methods: MNCs were collected from BM of the patients with CML in chronic phase by centrifugation in lymphocyte separation medium and cultured in liquid with IFN-a (200U/ml) or IFN-a (200U/ml) plus IL-6 (100 ng/ml). The growing cells were counted every day. The expression levels of b-actin, bcr-abl, bcl-2 and c-myc genes in the MNCs incubated for 24 h were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and relatively quantitative analysis of the amplified fragments by optical density scanning for the bands on gel. Results: The cell growth was markedly suppressed by IFN-a but the degree of cell-growth inhibition was slightly decreased by IL-6 on the basis of IFN-a effect. The expression of bcr-abl chimeric gene was intensely inhibited by IFN-a or IFN-a plus IL-6. The expression of bcl-2 gene was suppressed by either IFN-a or IFN-a plus IL-6, whereas that of c-myc gene was also inhibited by IFN-a but strongly elevated by IL-6 on the basis of IFN-a action. Conclusions: Both IFN-a and IFN-a plus IL-6 can inhibit the expression of anti-apoptosis gene such as bcr-abl and bcl-2 and regulate the expression of the cs.hn.cn gene related to cell proliferation and differentiation such as c-myc. Either IFN-a or IFN-a combined with IL-6 will serve as a trustful strategy of clinical treatment for CML.展开更多
The level of FcγR on peripheral blood mononuclear cells (PBMC) was determined by ABC-ELIsA in 43 patients with primary hepatocellular carcinoma (PHC) and in 8 patients with space-occupying benign lesion (BL) in liver...The level of FcγR on peripheral blood mononuclear cells (PBMC) was determined by ABC-ELIsA in 43 patients with primary hepatocellular carcinoma (PHC) and in 8 patients with space-occupying benign lesion (BL) in liver, twelve patients who received carcinomaectomy were followed up to detect their FcγR.Nine PBMC samples from PHC were treated with rIFNγ-The results showed that: 1. the level of FcγR in PHC patients was significantly lower than that in BL and In normal control; 2. the expression of FcγR was enhanced in postoperated PHC patients; and 3. the production of IFNγ in PHC patients was defective. The level of FcγR on PBMC was increased obviously in PHC after PBMC being treated with rIFNγ but was still lower than that in control. These results suggested; (1) FcyR detection is helpful tor distinguishing malignant liver lesion from benign; (2) the inhibited expression of FcγR in PHC could be related to some factors secreted by cancer tissues and also to low level of IFNγ; and (3) IFNγstimulates the expression of FcyR in vitro as one of the anti-tumor mechanism forIFNγ.展开更多
基金supported by Central Public-interest Scientific Institution Basal Research Fund(CATAS-Nos.1630152023007,1630152023011,1630152023012,1630152023013)the National Natural Science Foundation of China(Grant No.32071805).
文摘Coconut(Cocos nucifera L.),a major oil and fruit crop of the Arecaceae family,is extensively cultivated across the Asia—Pacific region.Despite its agricultural importance,genome assembly in coconut remains challenging due to its large genome size and high proportion of repetitive sequences.Allele-specific expression(ASE)plays a key role in regulating plant development and evolution,yet research on ASE in coconut is limited(Shao et al.,2019;Li et al.,2021;Zhang et al.,2021;Hu et al.,2022).Among phenotypic traits,fruit color is especially important as an indicator of maturity,guiding harvest timing and post-harvest processes(Kapoor et al.,2022).While prior studies have explored various coconut traits such as salt tolerance,fiber content,and plant height(Wang et al.,2021;Yang et al.,2021),investigations into ASE and fruit color remain scarce.
文摘In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl 2) in peripheral blood lymphocytes (PBLC) and the level of IFN γ and IL 4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl 2 in PBLC and the level of IFN γ and IL 4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl 2 was significantly lower (both P< 0.01). The level of IFN γ in serum of CA was significantly lower than in the normal control group ( P< 0.01), but IL 4 was significantly lower (both P< 0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN γ in serum of patients with CA, but had a positive correlation with the level of IL 4; The expression of Bcl 2 had a positive correlation with the level of IFN γ, but had a negative correlation with the level of IL 4. All the correlation coefficients had significant differerce by t test ( P< 0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the THl associated cytokines (eg: IFN γ) and the increased secretion of the TH2 asssociated cytokines (eg: IL 4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.
文摘Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the types of IFN, which suppresses proliferation and induces apoptosis in some cell types while inhibiting apoptosis in others. The aim of this study was to explore the effect of IFNα-2a on Fas expression and the apoptosis rate of peripheral blood cytotoxic T cells (CTLs) in patients with hepatitis B. METHODS:Peripheral blood mononuclear cells were isolated from 26 patients with hepatitis B including 16 patients with chronic hepatitis B and 10 patients with chronic severe hepatitis B. Fas expression and apoptosis rate of CTLs were analyzed with flow cytometry before and after IFNα-2a treatment. RESULTS:Before IFNα-2a treatment, Fas expression and apoptosis rate of CTLs from patients with chronic hepatitis B were significantly higher than those from patients with chronic severe hepatitis B and healthy controls respectively. No significant difference was observed between Fas expression and apoptosis rate of CTLs from patients with chronic severe hepatitis B and healthy controls. After IFNα-2a treatment,Fas expression and apoptosis rate of CTLs from different groups were compared with those before IFNα-2a treatment, showing no significant difference despite alternation of different degree. CONCLUSIONS:Activation induced cell death (AICD) exists in peripheral blood CTLs from patients with hepatitis B. No effect of IFNα-2a exerts on Fas expression and apoptosis rate of Fas in patients with hepatitis B.
基金This work was supported by a grant from the Scientific Research Foundation of Ministry of Public Health of PR China (No. 96-1-204).
文摘Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon cancer cell line CCL229. Methods: Using the molecular and biochemical techniques and electron microscopy to analyze the expression of iNOS, production of NO and growth characteristics of human colon cancer cells. Results: cytokine treatment can induce expression of the iNOS gene and production of nitric oxide was significantly higher after treatment of CCL229 cells with TNF-αor IFN-γ. Treatment with either cytokine or a combination of both significantly increased levels of Malondialdehyde (MDA) over control. Furthermore, cytokine treatment increased the proliferation inhibition rate as assessed in vitro and decreased the cell proliferation index on flow cytometry. Electron microscopy showed that cells treated with cytokines had fewer pseudopodia or cell processes than control cells and that cytokine treated cells had dilatation of the mitochondria and endoplasmic reticulum and dilated vesicular or tubular cisternae. Conclusion: Our findings indicate that TNF-α and IFN-γ induce the expression of iNOS gene in CCL229 cells, which increases the production of nitric oxide, inhibits proliferation, causes lipid peroxidation, and results in ultrastructural changes.
基金This study was supported by research grant from National Natural Science Foundation of China (No.30572446), research grants from Modern Biology & Pharmacy Foundation of ShanghaiScience Committee (No.02D219115) and Fudan University (985 Program).
文摘Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and cultured in vitro, and the expression of IL-10R1 and IL-10R2 gene was analyzed after cells had been treated with TH2-type cytokines IL-10 and TH1-type cytokines IFN-γ within 60 rain with semiquantitative reverse transcriptase-PCR, then the influence of IL-10 and IFN-γ on expression of IL-10R protein was examined by first trimester DSC using flow cytometry. In addition, the vitality of DSC was detected by MTT. Results IL-10R1 mRNA levels of DSC treated with IL-10 （10 ng/ml） reached the peak level within 15 rain, and were significantly lower at 30 rain, then were not detected at 45 min. The expression of IL-10R1 were induced to moderate level by IFN-γ（10 ng/ml） within 30 rain, and reduced to undetected levels at 60 min. There was no significant difference of IL-10R2 expression （P〉0.05） between treated and not with the abovementioned cytokines. The IL-10R protein expression and vitality of DSC were significantly enhanced by IL-10 （10 ng/ml） and IFN-γ （10 ng/ml） which treated DSC 48 h （P〈0.05）. Coneclusion IL-10 and IFN-γ may play an important role of biologic function in early pregnancy by influencing IL-10R expression of DSC.
基金Six Talent Peaks Project in Jiangsu Province(NY023)Horizontal Cooperation Project of Yangzhou Goo Sing Agriculture and Animal Husbandry Science and Technology Co.,Ltd.(00010114012,NSFPT201510)Special Fund of Jiangsu Huanenghui Medical Equipment Cytotoxicity Test(NSFPT201512)
文摘Because yeast codon preferred to synthesize the gene sequence of ChIFN-α mature peptide, Pichia pastoris expression system was connected into vector pPICgK, to construct recombinant plasmid pPIC9K-α. After digestion and linearization, the recombinant vector was cloned into P. pastor/s GS115, high copy transformant and its methanol utilization type were screened, and its expression conditions were optimized. The results showed that ChlFN-α successfully expressed in P. pastor/s, the expression amount of recombinant protein was the largest after induced expression in 0.5 % methanol containing 2% acid hydrolyzed casein at 28 ℃ for 72 h, and its antiviral activity was the highest.
文摘In ord,r to study the relatiouship between the expression or PML and the antiproliferative effect or interferon (IFN), prostate cancer cells, PC-3, DU145 and LNCaP were cultured, andtreated with IFN-α. The cell growth. rates were measured by crystal violet assay, and the expressionof PML was detected by immunofluomcent staining with PML Polyclonal antibody. Our study indicated that the growth rates or PC-3 and DU145 were significantly inhibited by IFN-α, whereas nosuch effect can be found on LNCaP cell line. The immunofluorescent staining showed that both thestaining intensity and the number or the PML nuclear boles were significantly increased in PC-3 andDU145 cells, however, no significant changes in the PML nuclear body were found in the IFs-αtreated LNCaP cells. Our study demeastrated that the antiproliferative,effct of IFN in prostate cancer cells may be directly related to the induced expression of PML in response to its treatment. Ourfinding supports the hypothesis that PML may mediate some antiproliferative erfects of IFN.
基金supported by National Nature Science Foundation of China(30700841)
文摘Objective： This study investigated the effects of rh-TRAIL with or without chemotherapeutic drugs on the apoptosis of the osteosarcoma cell line, MG-63, and the influence of chemotherapeutic drugs on changes in the expression of DR5 and YinYang 1 （YY1） in MG- 63 cells. Methods： The effects of treatment with rh-TRAIL alone and/or chemotherapetic drugs on MG-63 cell growth inhibition and apoptosis were measured using the MTT assay, FACS analysis ofAunexin V labeled cells, and the mRNA changes of DR5 and YY1 were detected by RT-PCR. Results： Rh-TRAIL protein inhibited the growth of MG-63 cells, and this inhibition was increased by adriamycin and IFN-γ. Adriamycin and IFN-γ significantly facilitated the induction of the expression of DR5 and reduced the expression of YY1. Conclusion： The apoptosis-inducing effect of rh-TRAIL in MG-63 cells was enhanced by chemotherapeutic drugs.
基金Supported by the Technology Program of Guangdong Province, No. 2004B35001001
文摘AIM： To study the association between host immunity and hepatitis B virus （HBV） recurrence after liver transplantation. METHODS： Peripheral blood mononuclear cells （PBMC） were isolated from 40 patients with hepatitis B and underwent orthotopic liver transplantation （OLT） before and 2, 4, 8 wk after surgery. After being cultured in vitro for 72 h, the levels of INF-γ and TNF-α in culture supematants were detected with ELISA. At the same time, the quantities of HBV DNA in serum and PBMCs were measured by real time PCR. RESULTS： The levels of INF-γ and TNF-α in PBMC culture supernatants decreased before and 2, 4 wk after surgery in turns （INF-γ 155.52±72.32 ng/L vs 14.76±9.88 ng/L vs 13.22±10.35 ng/L, F= 6.946, P= 0.027〈0.05; TNF-α 80.839±46.75 ng/L vs18.59±17.29 ng/L vs9.758±7.96 ng/L, F= 22.61, P = 0.0001〈0.05）. The levels of INF-γ and TNF-α were higher in groups with phytohemagglutinin （PHA） than in those without PHA before surgery. However, the difference disappeared following OLT. Furthermore, INF-γ and TNF-α could not be detected in most patients at wk 4 and none at wk 8 after OLT. The HBV detection rate and virus load in PBMC before and 2, 4 wk after surgery were fluctuated （HBV detected rate： 51.4%, 13.3%, 50% respectively; HBV DNA： 3.55±0.674 log（10） copies/mL vs 3.00±0.329 log（10） copies/mL vs 4.608±1.344 log（10） copies/mL, F= 7.582, P= 0.002〈0.05）. HBV DNA in serum was 4.48±1.463 log（10） copies/mL before surgery and 〈 10^3 copies/mL after OLT except for one with 5.72×10^6 copies/mL 4 wk after OLT who was diagnosed as HBV recurrence. The levels of INF-γ and TNF-α were lower in patients with a high HBV load than in those with a low HBV load （HBV DNA detected/undetected in PBMCs： IFN-γ 138.08±72.44 ng/L vs 164.24±72.07 ng/L, t = 1.065, P= 0.297〉0.05, TNF-α 80.75±47.30 ng/L vs 74.10±49.70 ng/L, t= 0.407, P= 0.686〉 0.05; HBV DNA positive/negative： IFN-γ 136.77±70.04 ng/L vs 175.27±71.50 ng/L, t= 1.702, P= 0.097〉0.05; TNF-α 75.37±43.02 ng/L vs 81.53±52.46 ng/L, t = 0.402, P = 0.690〉0.05）. CONCLUSION： The yielding of INF-γ and TNF-α from PBMCs is inhibited significantly by immunosuppressive agents following OLT with HBV load increased, indicating that the impaired immunity of host is associated with HBV recurrence after OLT.
基金the National Natural Science Foundation of China !(No. 39570805).
文摘Objective:To investigate the effects of interferon-a (IFN-a) and IFN-a combined with interleukin-6 (IL-6) on growth and expression of bcr-abl, bcl-2 and c--myc genes in the mononuclear cells (MNCs) from bone marrow (BM) of patients with chronic myelogenous leukemia (CML). Methods: MNCs were collected from BM of the patients with CML in chronic phase by centrifugation in lymphocyte separation medium and cultured in liquid with IFN-a (200U/ml) or IFN-a (200U/ml) plus IL-6 (100 ng/ml). The growing cells were counted every day. The expression levels of b-actin, bcr-abl, bcl-2 and c-myc genes in the MNCs incubated for 24 h were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and relatively quantitative analysis of the amplified fragments by optical density scanning for the bands on gel. Results: The cell growth was markedly suppressed by IFN-a but the degree of cell-growth inhibition was slightly decreased by IL-6 on the basis of IFN-a effect. The expression of bcr-abl chimeric gene was intensely inhibited by IFN-a or IFN-a plus IL-6. The expression of bcl-2 gene was suppressed by either IFN-a or IFN-a plus IL-6, whereas that of c-myc gene was also inhibited by IFN-a but strongly elevated by IL-6 on the basis of IFN-a action. Conclusions: Both IFN-a and IFN-a plus IL-6 can inhibit the expression of anti-apoptosis gene such as bcr-abl and bcl-2 and regulate the expression of the cs.hn.cn gene related to cell proliferation and differentiation such as c-myc. Either IFN-a or IFN-a combined with IL-6 will serve as a trustful strategy of clinical treatment for CML.
文摘The level of FcγR on peripheral blood mononuclear cells (PBMC) was determined by ABC-ELIsA in 43 patients with primary hepatocellular carcinoma (PHC) and in 8 patients with space-occupying benign lesion (BL) in liver, twelve patients who received carcinomaectomy were followed up to detect their FcγR.Nine PBMC samples from PHC were treated with rIFNγ-The results showed that: 1. the level of FcγR in PHC patients was significantly lower than that in BL and In normal control; 2. the expression of FcγR was enhanced in postoperated PHC patients; and 3. the production of IFNγ in PHC patients was defective. The level of FcγR on PBMC was increased obviously in PHC after PBMC being treated with rIFNγ but was still lower than that in control. These results suggested; (1) FcyR detection is helpful tor distinguishing malignant liver lesion from benign; (2) the inhibited expression of FcγR in PHC could be related to some factors secreted by cancer tissues and also to low level of IFNγ; and (3) IFNγstimulates the expression of FcyR in vitro as one of the anti-tumor mechanism forIFNγ.
