[Objective] This study aimed to investigate the changes of the transcriptional levels of molecules associated with endogenous antigen processing and presenta- tion in porcine skin-derived dendritic cells infected with...[Objective] This study aimed to investigate the changes of the transcriptional levels of molecules associated with endogenous antigen processing and presenta- tion in porcine skin-derived dendritic cells infected with PCV2 in vivo. [Method] Healthy 40-day-old Landrace piglets were infected with porcine circovirus type 2 (PCV2) and euthanized on the 34, 7rd, 14th, 21st and 35th d post inoculation (DPI). The porcine skin-derived dendritic cells (DCs) were collected to analyze the transcrip- tional levels of molecules (LMP7, UBP, MHC-I, calreticulin) associated with endogenous antigen processing and presentation by using real-time fluorescent quantitative PCR (real-time FQ-PCR). [Result] The results showed that the level of LMP7 mR- NAs was reduced significantly on the 3DPI (P〈0.05); the level of UBP mRNAs was consistently up-regulated, which increased significantly on the 21DPI and 35DPI (P〈 0.05); the level of MHC-I mRNAs was significantly down-regulated on the 7DPI (P〈 0.05); the level of calreticulin mRNAs was up-regulated slightly without significant dif- ference. [Conclusion] PCV2 can inhibit the endogenous antigen processing and presentation ability of porcine skin-derived DCs at early stages of infection.展开更多
AIM: To investigate a novel DNA vaccination based upon expression of the HBV e antigen fused to a heat shock protein (HSP) as a strategy to enhance DNA vaccine potency.METHODS: A pCMV-HBeAg-HSP DNA vaccine and a c...AIM: To investigate a novel DNA vaccination based upon expression of the HBV e antigen fused to a heat shock protein (HSP) as a strategy to enhance DNA vaccine potency.METHODS: A pCMV-HBeAg-HSP DNA vaccine and a control DNA vaccine were generated. Mice were immunized with these different construct. Immune responses were measured 2 wk after a second immunization by a T cell response assay, CTL cytotoxicity assay, and an antibody assay in C57BL/6 and BALB/c mice. CT26-HBeAg tumor cell challenge test in vivo was Performed in BALB/c mice to monitor anti-tumor immune responses.RESULTS: In the mice immunized with pCMV-HBe-HSP DNA, superior CTL activity to target HBV-positive target cells was observed in comparison with mice immunized with pCMV-HBeAg (44% ± 5% vs 30% ± 6% in E: T 〉 50:1, P 〈 0,05), ELISPOT assays showed a stronger T-cell response from mice immunized with pCMV-HBe- HSP than that from pCMV-HBeAg immunized animals when stimulated either with MHC class I or class Ⅱ epitopes derived from HBeAg (74% ± 9% vs 31% ± 6%, P 〈 0.01). ELISA assays revealed an enhanced HBeAg antibody response from mice immunized with pCMV- HBe-HSP than from those immunized with pCMV-HBeAg. The lowest tumor incidence and the slowest tumor growth were observed in mice immunized with pCMV- HBe-HSP when challenged with CT26-HBeAg.CONCLUSION: The results of this study demonstrate a broad enhancement of antigen-specific CD4^+ helper,CD8^+ cytotoxic T-cell, and B-cell responses by a novel DNA vaccination strategy. They also proved a stronger antigen-specific immune memory, which may be superior to currently described HBV DNA vaccination strategies for the treatment of chronic HBV infection.展开更多
Coxsackie virus A16(CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease(HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and ...Coxsackie virus A16(CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease(HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews(Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection.展开更多
AIM: To study the induction of T cellular immune responses in BALB/c mice immunized with uric acid and dendritic cells (DCs) pulsed with hepatitis B virus surface antigen (HBsAg). METHODS: DCs were generated fro...AIM: To study the induction of T cellular immune responses in BALB/c mice immunized with uric acid and dendritic cells (DCs) pulsed with hepatitis B virus surface antigen (HBsAg). METHODS: DCs were generated from bone-marrow cells of BABL/c mice, and then pulsed or unpulsed with HBsAg protein (HBsAg-pulsed-DCs or unpulsed-DCs) in vitro. BABL/c mice were immunized with HBsAg-pulsed- DCs (1 × 10^6) and uric acid, injected through the tail vein of each mouse. The mice in control groups were immunized with HBsAg-pulsed-DCs alone, unpulsed- DCs alone or 200 μg uric acid alone or PBS alone. The immunization was repeated 7 d later. Cytotoxic T lymphocytes (CTLs) in vivo were determined by the CFSE labeled spleen lysis assay. Spleen cells or spleen T cells were isolated, and re-stimulated in vitro with HBsAg for 120 h or 72 h. Production of IFN-γ, and IL-4 secreted by spleen cells were determined by ELISA method; proliferation of spleen T cells were detected by flow cytometry. RESULTS: The cytotoxicities of HBsAg-specific-CTLs, generated after immunization of HBsAg-pulsed-DCs and uric acid, were 68.63% ±11.32% and significantly stronger than that in the control groups (P 〈 0.01). Compared with control groups, in mice treated with uric acid and HBsAg-pulsed-DCs, the spleen T cell proliferation to HBsAg re-stimulation was stronger (1.34 ± 0.093 vs 1.081±0.028, P 〈 0.01), the level of IFN-t, secreted by splenocytes was higher (266.575 ± 51.323 vs 135.223 ±32.563, P 〈 0.01) , and IL-4 level wasower (22.385 ± 2.252 vs 40.598 ± 4.218, P 〈 0.01). CONCLUSION: Uric acid can strongly enhance T cell immune responses induced by HBsAg-pulsed-DCs vaccine. Uric acid may serve as an effective adjuvant of DC vaccine against HBV infection.展开更多
Recent developments in tumor immunology and biotechnology have made cancer gene therapy and immunotherapy feasible. The current efforts for cancer gene therapy mainly focus on using immunogenes, chemogenes and tumor s...Recent developments in tumor immunology and biotechnology have made cancer gene therapy and immunotherapy feasible. The current efforts for cancer gene therapy mainly focus on using immunogenes, chemogenes and tumor suppressor genes. Central to all these therapies is the development of efficient vectors for gene therapy. By far, adenovirus (AdV)-mediated gene therapy is one of the most promising approaches, as has confirmed by studies relating to animal tumor models and clinical trials. Dendritic cells (DCs) are highly efficient, specialized antigen-presenting cells, and DC- based tumor vaccines are regarded as having much potential in cancer immunotherapy. Vaccination with DCs pulsed with tumor peptides, lysates, or RNA, or loaded with apoptotic/necrotic tumor cells, or engineered to express certain cytokines or chemokines could induce significant antitumor cytotoxic T lymphocyte (CTL) responses and antitumor immunity. Although both AdV-mediated gene therapy and DC vaccine can both stimulate antitumor immune responses, their therapeutic efficiency has been limited to generation of prophylactic antitumor immunity against re-challenge with the parental tumor cells or to growth inhibition of small tumors. However, this approach has been unsuccessful in combating well-established tumors in animal models. Therefore, a major strategic goal of current cancer immunotherapy has become the development of novel therapeutic strategies that can combat well-established tumors, thus resembling real clinical practice since a good proportion of cancer patients generally present with significant disease. In this paper, we review the recent progress in AdV-mediated cancer gene therapy and DC-based cancer vaccines, and discuss combined immunotherapy including gene therapy and DC vaccines. We underscore the fact that combined therapy may have some advantages in combating well-established tumors vis-a-vis either modality administered as a monotherapy.展开更多
Aims: To screen for the predominant bacteria strains distributed in clean rooms and to analyze their phylogenetic relationships. Methods and Results: The bacteria distributed in air, surfaces and personnel in clean ro...Aims: To screen for the predominant bacteria strains distributed in clean rooms and to analyze their phylogenetic relationships. Methods and Results: The bacteria distributed in air, surfaces and personnel in clean rooms were routinely moni-tored using agar plates. Five isolates frequently isolated from the clean rooms of an aseptic pharmaceutical production workshop were selected based on their colony and cell morphology characteristics. Their physiological and biochemical properties, as well as partial 16S rDNA sequences, were analyzed. Results showed that all the five isolates belong to Gram positive bacteria, of which three were Staphylococcus, one Microbacterium and one Bacillus species. Sensitivity tests for these bacteria isolates to 3 disin-fectants showed that isolate F03 was obtuse, and had low susceptivity to UV irradiation, while isolates F02, F01 and F04 were not sensitive to phenol treatment. Isolates F04, F01 and F05 were resistant to chlorhexidine gluconate. Conclusion: Bacteria widely distributed in clean rooms are mainly a group of Gram positive strains, showing high resistance to selected disinfectants. Sig-nificance and impact of the study: Clean rooms are essential in aseptic pharmaceutical and food production. Screening bacteria isolates and identifying them is part of good manufacturing practices, and will aid in finding a more effective disinfection method.展开更多
The human hepatitis B virus (HBV) and the duck hepatitis B virus (DHBV) share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and th...The human hepatitis B virus (HBV) and the duck hepatitis B virus (DHBV) share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and the coding open reading frames (ORFs) overlap extensively. In addition, the genomic and structural organization, as well as replication and biological characteristics, are very similar in both viruses. Host of the key features of hepadnaviral infection were first discovered in the DHBV model system and subsequently confirmed for HBV. There are, however, several differences between human HBV and DHBV. This review will focus on the molecular and cellular biology, evolution, and host adaptation of the avian hepatitis B viruses with particular emphasis on DHBV as a model system.展开更多
The antiviral activity in vitro and in vivo and the effect of the immune system of two fucoidan fractions with low molecular weight and different sulfate content from Laminaria japonica(LMW fucoidans) were investigate...The antiviral activity in vitro and in vivo and the effect of the immune system of two fucoidan fractions with low molecular weight and different sulfate content from Laminaria japonica(LMW fucoidans) were investigated in order to examine the possible mechanism. In vitro, I-type influenza virus, adenovirus and Parainfluenza virus I were used to infect Hep-2, Hela and MDCK cells, respectively. And 50% tissue culture infective dose was calculated to detect the antiviral activity of two LMW fucoidans. The results indicated that compared with the control group, 2 kinds of LMW fucoidans had remarkable antiviral activity in vitro in middle and high doses, while at low doses, the antiviral activity of 2 kinds of LMW fucoidans was not statistically different from that in the blank control group. And there was no statistically difference between two LMW fucoidans in antiviral activity. In vivo, LMW fucoidans could prolong the survival time of virus-infected mice, and could improve the lung index of virus-infected mice significantly, which have statistical differences with the control group significantly(p < 0.01). However, the survival time of the two LMW fucoidans was not statistically significant(p > 0.05). In this study, it was shown that both of two LMW fucoidans(LF1, LF2) could increase the thymus index, spleen index, phagocytic index, phagocytosis coefficient and half hemolysin value in middle and high doses, which suggested that LMW fucoidans could play an antiviral role by improving the quality of immune organs, improving immune cell phagocytosis and humoral immunity.展开更多
AIM: To investigate the in-vitro activation of cytotoxic T lymphocytes (CTLs) by fusion of mouse hepatocellular carcinoma (HCC) ceils and lymphotactin gene-modified dendritic cells (DCs). METHODS: Lymphotactin...AIM: To investigate the in-vitro activation of cytotoxic T lymphocytes (CTLs) by fusion of mouse hepatocellular carcinoma (HCC) ceils and lymphotactin gene-modified dendritic cells (DCs). METHODS: Lymphotactin gene modified DCs (DCLptn) were prepared by lymphotactin recombinant adenovirus transduction of mature DCs which differentiated from mouse bone marrow cells by stimulation with granulocyte/macrophage colony-stimulating factor (GM- CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-α). DCLptn and H22 fusion was prepared using 50% PEG. Lymphotactin gene and protein expression levels were measured by RT-PCR and ELISA, respectively. Lymphotactin chemotactic responses were examined by in-vitro chemotaxis assay. In-vitro activation of CTl_s by DCLptn/H22 fusion was measured by detecting CD25 expression and cytokine production after autologous T cell stimulation. Cytotoxic function of activated T lymphocytes stimulated with DCLptn/H22 cells was determined by LDH cytotoxicity assay. RESULTS: Lymphotactin gene could be efficiently transduced to DCs by adenovirus vector and showed an effective biological activity. After fusion, the hybrid DCLptn/H22 cells acquired the phenotypes of both DCLptn and H22 cells. In T cell proliferation assay, flow cytometry showed a very high CD25 expression, and cytokine release assay showed a significantly higher concentration of IFN-α, and IL-2 in DCLptn/H22 group than in DCLptn, DCLptn+H22, DC/H22 or H22 groups. Cytotoxicity assay revealed that T cells derived from DCLptn/H22 group had much higher anti-tumor activity than those derived from DCLptn, H22, DCLptn + H22, DC/H22 groups. CONCLUSION: Lymphotactin gene-modified dendritoma induces T-cell proliferation and strong CTL reaction against allogenic HCC cells. Immunization-engineered fusion hybrid vaccine is an attractive strategy in prevention and treatment of HCC metastases.展开更多
Hepatitis B virus (HBV) infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of...Hepatitis B virus (HBV) infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of chronic HBV carriers suffer from progressive liver diseases. In addition, all HBV carriers are permanent source of this virus. There is no curative therapy for chronic HBV carriers. Antiviral drugs are recommended for about 10% patients, however, these drugs are costly, have limited efficacy, and possess considerable side effects. Recent studies have shown that immune responses of the host to the HBV are critically involved at every stage of chronic HBV infection: (1) These influence acquisition of chronic HBV carrier state, (2) They are important in the context of liver damages, (3) Recovery from chronic HBV-related liver diseases is dependent on nature and extent of HBV-specific immune responses. However, induction of adequate levels of HBV-specific immune responses in chronic HBV carriers is difficult. During the last one decade, hepatitis B vaccine has been administered to chronic HBV carriers as a therapeutic approach (vaccine therapy). The present regimen of vaccine therapy is safe and cheap, but not so effective. A dendritic cell-based therapeutic vaccine has recently been developed for treating chronic HBV infection. In this review, we will discuss about the concept, scientific logics, strategies and techniques of development of HBV- specific immune therapies including vaccine therapy and dendritic cell-based vaccine therapy for treating chronic HBV infection.展开更多
It was generally considered that contamination of the gel type polystyrenestrong basic anion exchange resin by organic matter in natural water is the result ofion exchange and Van der waal's adsorption on it. On t...It was generally considered that contamination of the gel type polystyrenestrong basic anion exchange resin by organic matter in natural water is the result ofion exchange and Van der waal's adsorption on it. On the basis of laboratory and industrial experiments, this paper confirmed that the interreaction between organicmatter and resin polymer matrix is primarily controled by a Van der waal's adsorption.展开更多
Due to the elevated ecological awareness nowadays the consumption of products of organic agriculture is increasing. Organic farming means excluding the use of synthetic inputs, such as synthetic fertilizers and pestic...Due to the elevated ecological awareness nowadays the consumption of products of organic agriculture is increasing. Organic farming means excluding the use of synthetic inputs, such as synthetic fertilizers and pesticides or genetically modified organisms, but there are only a few regulations regarding heavy metal concentrations in soils. Thus a not negligible uptake of metals from the soil where the apple trees are grown may occur. Furthermore inorganic copper compounds being traditional fertilizers for apple trees are not considered as synthetic fertilizers, thus they are still used in organic apple farming for soil or foliar application. Thus also apples produced by organic agriculture may contain toxic elements, such as cadmium, chromium, copper, and lead. The concentrations of these elements were determined in whole apples, as well as in the flesh and peel in order to estimate the possible risk for human health. Prior to analysis using ICP-AES samples underwent a microwave assisted digestion. The LODs obtained are below the recommended maximum levels in vegetables/fruits by WHO. In all samples no Cu, Cr, and Pb could be detected. Only Ca was found in the peel of about a third of samples investigated in concentrations between LOD and LOQ (1-3 μg/g).展开更多
The effects of acid deposition on pine forest ecosystems in Longli of Guizhou Province, southwestern China are studied using indoor experiments and model simulations. Indoor experiments are designed to explore the alu...The effects of acid deposition on pine forest ecosystems in Longli of Guizhou Province, southwestern China are studied using indoor experiments and model simulations. Indoor experiments are designed to explore the aluminum toxicity on pine seedlings, and the long-term soil acidification model (LTSAM) and a terrestrial biogeochemistry model (CENTURY) are used to simulate the influences of acid deposition on pine forest ecosystems. The indoor experiment results of aluminum toxicishow that aluminum ions in solution limit plant growth and acid deposition enhances this effect by facilitating the release of aluminum ions from the soil. Pine seedling bio- mass and root elongation decrease as the aluminum concentration increases. The results of model simulations show that the soil chemis- try varies significantly with different changes in acid deposition. When the acid deposition increases, the pH value in the soil solution decreases and the soil A13+ concentration increases. The increased acid deposition also has negative impacts on the forest ecosystem, i.e., decreases plant biomass, net primary productivity (NPP) and net C02 uptake. As a result, the soil organic carbon (SOC) decreases be- cause of the limited supply of decomposition material. Thus acid deposition need be reduced to help protect the forest ecosystems.展开更多
AIM: To explore dendritic cells (DCs) multiple functions in immune modulation. METHODS: We used bone-marrow derived dendritic cells from BALB/c mice pulsed with pseudo particles from the hepatitis C virus to vaccinate...AIM: To explore dendritic cells (DCs) multiple functions in immune modulation. METHODS: We used bone-marrow derived dendritic cells from BALB/c mice pulsed with pseudo particles from the hepatitis C virus to vaccinate naive BALB/c mice. Hepatitis C virus (HCV) pseudo particles consist of the genotype 1b derived envelope proteins E1 and E2, covering a non-HCV core structure. Thus, not a single epitope, but the whole "viral surface" induces immunogenicity. For vaccination, mature and activated DC were injected subcutaneously twice. RESULTS: Humoral and cellular immune responses measured by enzyme-linked immunosorbent assay and interferon-gamma enzyme-linked immunosorbent spot test showed antibody production as well as T-cellsdirected against HCV. Furthermore, T-cell responses confi rmed two highly immunogenic regions in E1 and E2 outside the hypervariable region 1. CONCLUSION: Our results indicate dendritic cells as a promising vaccination model for HCV infection that should be evaluated further.展开更多
The deoiled Jatropha curcas meal by hexane extraction was detoxified phorbol esters by two different methods. These two methods were alkali in methanol and only ethanol washing. After both treatments, the PEs (phorbo...The deoiled Jatropha curcas meal by hexane extraction was detoxified phorbol esters by two different methods. These two methods were alkali in methanol and only ethanol washing. After both treatments, the PEs (phorbol esters) was decreased by 100%. The crude protein in detoxified meal of alkali in methanol washing was less amount than only ethanol washing. The result showed that treatment by only ethanol washing was a promising way to detoxify deoiled Jatropha curcas meal for animal feeds in industrial scale.展开更多
Invasions by exotic insects and diseases are one of the most important threats to the stability and productivity of forest ecosystems around the world. Lymantria dispar (Lepidoptera: Lymantriidae) in Iran was obser...Invasions by exotic insects and diseases are one of the most important threats to the stability and productivity of forest ecosystems around the world. Lymantria dispar (Lepidoptera: Lymantriidae) in Iran was observed for the first time in 1937 by Jalal Afshar in oak forests of the Guilan state region. Since its introduction, gypsy moth has spread to the northern, western and south-western regions of Iran. The largest outbreaks of gypsy moth occurred in the Talesh Oak forests of the Hyrcanian zone in 1975. The gypsy moth is a highly polyphagous folivore species that feeds on over 300 species of woody plants. Among its most preferred hosts are oaks and aspens. Some trees are resistant to the gypsy moth including honey locust, black locust, silver maple, green ash, dogwood, sycamore, horse chestnut, firs, and tulip trees. Defoliation during severe outbreaks alters the appearance of forest habitats and leads to tree mortality and changes in stand composition. This study was conducted during 2008 and 2009 in an effort to determine the primary host tree species that the gypsy moth infests in the Hyrcanian forests of lran. For that purpose all egg masses in the area were counted and conclusively significant differences in defoliation levels among tree species were found. The primary host tree species of gypsy moth in Iran was Persian ironweed, Parrotia persica.展开更多
A two-step method was developed to quantitatively assess the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis (Li) Humber, on the green peach aphid, Myzus persicae (Sulzer) .Firstly, a standard t...A two-step method was developed to quantitatively assess the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis (Li) Humber, on the green peach aphid, Myzus persicae (Sulzer) .Firstly, a standard time-dose-mortality relationship, established by modeling data from bioassay 1 at varying conidial dosages (0.4- 10.4 conidia/mm^2) of Z. anhuiensis F97028, was used to yield an estimate of expected mortality probability at a given dosage. Secondly, bioassay 2 was conducted by simultaneously exposing six ≤4-day-old nymphal colonies to a shower of Z. anhuiensis conidia at each of four dosages (resulting from exposures of 0.3 - 8.0 min) . Subsequently, the colonies were separately immersed in a 0.1% chlorothalonil solution for 0.5 min to disinfect all surviving conidia on the host integument from 1 - 12 h after exposure under temperature treatments of 15 and 20℃, respectively. The infection rate during a specific period from the end of the exposure to the immersion was then estimated as the ratio of the observed mortality over the expected mortality probability at a particular dosage. The results showed that the infection of M. persicae from Z. anhuiensis was highly rapid with little difference between aphid colonies maintained at 15 and 20℃ before being immersed in the fungicidal solution after exposure. The first 6-hour period after exposure was most crucial to successful infection of the fungus with the infection rate greatly depending on conidial dosages. It took ≤ 1 h to infect > 50% of the aphids at a dosage of > 1.5 conida/mm^2 and > 90% at > 50 conidia/mm^2 .展开更多
In this research paper, our main objective is to find out the meticulous role of activated dendritic cells (DCs) during the human immunodeficiency virus (HIV) infection process. DCs play a dual role by enhancing b...In this research paper, our main objective is to find out the meticulous role of activated dendritic cells (DCs) during the human immunodeficiency virus (HIV) infection process. DCs play a dual role by enhancing both HIV infection progression, as well as antiviral immune response. To explore the implications of these dual roles, we have formulated our mathematical model and analyzed the model by both analytical and numerical approaches. By using an impulsive differential equation, we have studied the effect of DC-based vaccination. Analytically we have determined the threshold value of drug dosage and dosing interval for optimum levels of injection. We have also investigated the effect of perfect adherence of drug dose on the immune cell count in extreme cases and observed that, systematic drug dose of the immune cells leads to its maximum level.展开更多
A new technique for mass rearing Clostera anachoreta(C.) was developed through a large numberof experiments, so the problem of mass produce of CaGV was solved. The process of CaGV was discussedin detail.
基金Supported by Natural Science Foundation of Beijing "Effect of porcine skin-derived dendritic cells on PCV infection" (6062006)Beijing Organization Department Project"Influence of PCV infection on bone marrow cell differentiation" (20061D0502100282)~~
文摘[Objective] This study aimed to investigate the changes of the transcriptional levels of molecules associated with endogenous antigen processing and presenta- tion in porcine skin-derived dendritic cells infected with PCV2 in vivo. [Method] Healthy 40-day-old Landrace piglets were infected with porcine circovirus type 2 (PCV2) and euthanized on the 34, 7rd, 14th, 21st and 35th d post inoculation (DPI). The porcine skin-derived dendritic cells (DCs) were collected to analyze the transcrip- tional levels of molecules (LMP7, UBP, MHC-I, calreticulin) associated with endogenous antigen processing and presentation by using real-time fluorescent quantitative PCR (real-time FQ-PCR). [Result] The results showed that the level of LMP7 mR- NAs was reduced significantly on the 3DPI (P〈0.05); the level of UBP mRNAs was consistently up-regulated, which increased significantly on the 21DPI and 35DPI (P〈 0.05); the level of MHC-I mRNAs was significantly down-regulated on the 7DPI (P〈 0.05); the level of calreticulin mRNAs was up-regulated slightly without significant dif- ference. [Conclusion] PCV2 can inhibit the endogenous antigen processing and presentation ability of porcine skin-derived DCs at early stages of infection.
文摘AIM: To investigate a novel DNA vaccination based upon expression of the HBV e antigen fused to a heat shock protein (HSP) as a strategy to enhance DNA vaccine potency.METHODS: A pCMV-HBeAg-HSP DNA vaccine and a control DNA vaccine were generated. Mice were immunized with these different construct. Immune responses were measured 2 wk after a second immunization by a T cell response assay, CTL cytotoxicity assay, and an antibody assay in C57BL/6 and BALB/c mice. CT26-HBeAg tumor cell challenge test in vivo was Performed in BALB/c mice to monitor anti-tumor immune responses.RESULTS: In the mice immunized with pCMV-HBe-HSP DNA, superior CTL activity to target HBV-positive target cells was observed in comparison with mice immunized with pCMV-HBeAg (44% ± 5% vs 30% ± 6% in E: T 〉 50:1, P 〈 0,05), ELISPOT assays showed a stronger T-cell response from mice immunized with pCMV-HBe- HSP than that from pCMV-HBeAg immunized animals when stimulated either with MHC class I or class Ⅱ epitopes derived from HBeAg (74% ± 9% vs 31% ± 6%, P 〈 0.01). ELISA assays revealed an enhanced HBeAg antibody response from mice immunized with pCMV- HBe-HSP than from those immunized with pCMV-HBeAg. The lowest tumor incidence and the slowest tumor growth were observed in mice immunized with pCMV- HBe-HSP when challenged with CT26-HBeAg.CONCLUSION: The results of this study demonstrate a broad enhancement of antigen-specific CD4^+ helper,CD8^+ cytotoxic T-cell, and B-cell responses by a novel DNA vaccination strategy. They also proved a stronger antigen-specific immune memory, which may be superior to currently described HBV DNA vaccination strategies for the treatment of chronic HBV infection.
基金supported by the National High-Tech R&D Program(2014ZX09102042)the National Natural Science Foundation of China(81373142)the Natural Science Foundation of Yunnan Province(2012ZA009)
文摘Coxsackie virus A16(CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease(HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews(Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection.
基金Supported by the National Natural Science Foundation of China, No. 30471533
文摘AIM: To study the induction of T cellular immune responses in BALB/c mice immunized with uric acid and dendritic cells (DCs) pulsed with hepatitis B virus surface antigen (HBsAg). METHODS: DCs were generated from bone-marrow cells of BABL/c mice, and then pulsed or unpulsed with HBsAg protein (HBsAg-pulsed-DCs or unpulsed-DCs) in vitro. BABL/c mice were immunized with HBsAg-pulsed- DCs (1 × 10^6) and uric acid, injected through the tail vein of each mouse. The mice in control groups were immunized with HBsAg-pulsed-DCs alone, unpulsed- DCs alone or 200 μg uric acid alone or PBS alone. The immunization was repeated 7 d later. Cytotoxic T lymphocytes (CTLs) in vivo were determined by the CFSE labeled spleen lysis assay. Spleen cells or spleen T cells were isolated, and re-stimulated in vitro with HBsAg for 120 h or 72 h. Production of IFN-γ, and IL-4 secreted by spleen cells were determined by ELISA method; proliferation of spleen T cells were detected by flow cytometry. RESULTS: The cytotoxicities of HBsAg-specific-CTLs, generated after immunization of HBsAg-pulsed-DCs and uric acid, were 68.63% ±11.32% and significantly stronger than that in the control groups (P 〈 0.01). Compared with control groups, in mice treated with uric acid and HBsAg-pulsed-DCs, the spleen T cell proliferation to HBsAg re-stimulation was stronger (1.34 ± 0.093 vs 1.081±0.028, P 〈 0.01), the level of IFN-t, secreted by splenocytes was higher (266.575 ± 51.323 vs 135.223 ±32.563, P 〈 0.01) , and IL-4 level wasower (22.385 ± 2.252 vs 40.598 ± 4.218, P 〈 0.01). CONCLUSION: Uric acid can strongly enhance T cell immune responses induced by HBsAg-pulsed-DCs vaccine. Uric acid may serve as an effective adjuvant of DC vaccine against HBV infection.
文摘Recent developments in tumor immunology and biotechnology have made cancer gene therapy and immunotherapy feasible. The current efforts for cancer gene therapy mainly focus on using immunogenes, chemogenes and tumor suppressor genes. Central to all these therapies is the development of efficient vectors for gene therapy. By far, adenovirus (AdV)-mediated gene therapy is one of the most promising approaches, as has confirmed by studies relating to animal tumor models and clinical trials. Dendritic cells (DCs) are highly efficient, specialized antigen-presenting cells, and DC- based tumor vaccines are regarded as having much potential in cancer immunotherapy. Vaccination with DCs pulsed with tumor peptides, lysates, or RNA, or loaded with apoptotic/necrotic tumor cells, or engineered to express certain cytokines or chemokines could induce significant antitumor cytotoxic T lymphocyte (CTL) responses and antitumor immunity. Although both AdV-mediated gene therapy and DC vaccine can both stimulate antitumor immune responses, their therapeutic efficiency has been limited to generation of prophylactic antitumor immunity against re-challenge with the parental tumor cells or to growth inhibition of small tumors. However, this approach has been unsuccessful in combating well-established tumors in animal models. Therefore, a major strategic goal of current cancer immunotherapy has become the development of novel therapeutic strategies that can combat well-established tumors, thus resembling real clinical practice since a good proportion of cancer patients generally present with significant disease. In this paper, we review the recent progress in AdV-mediated cancer gene therapy and DC-based cancer vaccines, and discuss combined immunotherapy including gene therapy and DC vaccines. We underscore the fact that combined therapy may have some advantages in combating well-established tumors vis-a-vis either modality administered as a monotherapy.
基金Project (No. 30371703) supported by the National Natural ScienceFoundation of China
文摘Aims: To screen for the predominant bacteria strains distributed in clean rooms and to analyze their phylogenetic relationships. Methods and Results: The bacteria distributed in air, surfaces and personnel in clean rooms were routinely moni-tored using agar plates. Five isolates frequently isolated from the clean rooms of an aseptic pharmaceutical production workshop were selected based on their colony and cell morphology characteristics. Their physiological and biochemical properties, as well as partial 16S rDNA sequences, were analyzed. Results showed that all the five isolates belong to Gram positive bacteria, of which three were Staphylococcus, one Microbacterium and one Bacillus species. Sensitivity tests for these bacteria isolates to 3 disin-fectants showed that isolate F03 was obtuse, and had low susceptivity to UV irradiation, while isolates F02, F01 and F04 were not sensitive to phenol treatment. Isolates F04, F01 and F05 were resistant to chlorhexidine gluconate. Conclusion: Bacteria widely distributed in clean rooms are mainly a group of Gram positive strains, showing high resistance to selected disinfectants. Sig-nificance and impact of the study: Clean rooms are essential in aseptic pharmaceutical and food production. Screening bacteria isolates and identifying them is part of good manufacturing practices, and will aid in finding a more effective disinfection method.
基金Supported by the Freie und Hansestadt Hamburg and the Bundesministcrium für Gesundheit und Soziale Sicherung grants from DFG and by the German Competence Network for Viral Hepatitis (Hop-Net), funded by the German Ministry of Education and Research (BMBF), Grant No. TFI3. IWe apologize to those authors whose work we could not cite directly due to space limitations. The authors are indebted to Claudia Franke (Heinrich-Pette-Institute, Hamburg, Germany) for providing the picture of core protein phosphorylation.
文摘The human hepatitis B virus (HBV) and the duck hepatitis B virus (DHBV) share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and the coding open reading frames (ORFs) overlap extensively. In addition, the genomic and structural organization, as well as replication and biological characteristics, are very similar in both viruses. Host of the key features of hepadnaviral infection were first discovered in the DHBV model system and subsequently confirmed for HBV. There are, however, several differences between human HBV and DHBV. This review will focus on the molecular and cellular biology, evolution, and host adaptation of the avian hepatitis B viruses with particular emphasis on DHBV as a model system.
基金supported by grants from the Technology Development Project (No. 201505022)the NSFC Shandong Joint Fund (No. U1406402)National Natural Science Foundation of China (No. 31371759)
文摘The antiviral activity in vitro and in vivo and the effect of the immune system of two fucoidan fractions with low molecular weight and different sulfate content from Laminaria japonica(LMW fucoidans) were investigated in order to examine the possible mechanism. In vitro, I-type influenza virus, adenovirus and Parainfluenza virus I were used to infect Hep-2, Hela and MDCK cells, respectively. And 50% tissue culture infective dose was calculated to detect the antiviral activity of two LMW fucoidans. The results indicated that compared with the control group, 2 kinds of LMW fucoidans had remarkable antiviral activity in vitro in middle and high doses, while at low doses, the antiviral activity of 2 kinds of LMW fucoidans was not statistically different from that in the blank control group. And there was no statistically difference between two LMW fucoidans in antiviral activity. In vivo, LMW fucoidans could prolong the survival time of virus-infected mice, and could improve the lung index of virus-infected mice significantly, which have statistical differences with the control group significantly(p < 0.01). However, the survival time of the two LMW fucoidans was not statistically significant(p > 0.05). In this study, it was shown that both of two LMW fucoidans(LF1, LF2) could increase the thymus index, spleen index, phagocytic index, phagocytosis coefficient and half hemolysin value in middle and high doses, which suggested that LMW fucoidans could play an antiviral role by improving the quality of immune organs, improving immune cell phagocytosis and humoral immunity.
基金Supported by the Science & Technology Foundation for Academicians of Zhejiang Province, China, No. 203201513
文摘AIM: To investigate the in-vitro activation of cytotoxic T lymphocytes (CTLs) by fusion of mouse hepatocellular carcinoma (HCC) ceils and lymphotactin gene-modified dendritic cells (DCs). METHODS: Lymphotactin gene modified DCs (DCLptn) were prepared by lymphotactin recombinant adenovirus transduction of mature DCs which differentiated from mouse bone marrow cells by stimulation with granulocyte/macrophage colony-stimulating factor (GM- CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-α). DCLptn and H22 fusion was prepared using 50% PEG. Lymphotactin gene and protein expression levels were measured by RT-PCR and ELISA, respectively. Lymphotactin chemotactic responses were examined by in-vitro chemotaxis assay. In-vitro activation of CTl_s by DCLptn/H22 fusion was measured by detecting CD25 expression and cytokine production after autologous T cell stimulation. Cytotoxic function of activated T lymphocytes stimulated with DCLptn/H22 cells was determined by LDH cytotoxicity assay. RESULTS: Lymphotactin gene could be efficiently transduced to DCs by adenovirus vector and showed an effective biological activity. After fusion, the hybrid DCLptn/H22 cells acquired the phenotypes of both DCLptn and H22 cells. In T cell proliferation assay, flow cytometry showed a very high CD25 expression, and cytokine release assay showed a significantly higher concentration of IFN-α, and IL-2 in DCLptn/H22 group than in DCLptn, DCLptn+H22, DC/H22 or H22 groups. Cytotoxicity assay revealed that T cells derived from DCLptn/H22 group had much higher anti-tumor activity than those derived from DCLptn, H22, DCLptn + H22, DC/H22 groups. CONCLUSION: Lymphotactin gene-modified dendritoma induces T-cell proliferation and strong CTL reaction against allogenic HCC cells. Immunization-engineered fusion hybrid vaccine is an attractive strategy in prevention and treatment of HCC metastases.
基金Supported by the Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan to Sk. Md. Fazle Akbar, No. C17590651 and Morikazu Onji, No. C17590652
文摘Hepatitis B virus (HBV) infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of chronic HBV carriers suffer from progressive liver diseases. In addition, all HBV carriers are permanent source of this virus. There is no curative therapy for chronic HBV carriers. Antiviral drugs are recommended for about 10% patients, however, these drugs are costly, have limited efficacy, and possess considerable side effects. Recent studies have shown that immune responses of the host to the HBV are critically involved at every stage of chronic HBV infection: (1) These influence acquisition of chronic HBV carrier state, (2) They are important in the context of liver damages, (3) Recovery from chronic HBV-related liver diseases is dependent on nature and extent of HBV-specific immune responses. However, induction of adequate levels of HBV-specific immune responses in chronic HBV carriers is difficult. During the last one decade, hepatitis B vaccine has been administered to chronic HBV carriers as a therapeutic approach (vaccine therapy). The present regimen of vaccine therapy is safe and cheap, but not so effective. A dendritic cell-based therapeutic vaccine has recently been developed for treating chronic HBV infection. In this review, we will discuss about the concept, scientific logics, strategies and techniques of development of HBV- specific immune therapies including vaccine therapy and dendritic cell-based vaccine therapy for treating chronic HBV infection.
文摘It was generally considered that contamination of the gel type polystyrenestrong basic anion exchange resin by organic matter in natural water is the result ofion exchange and Van der waal's adsorption on it. On the basis of laboratory and industrial experiments, this paper confirmed that the interreaction between organicmatter and resin polymer matrix is primarily controled by a Van der waal's adsorption.
文摘Due to the elevated ecological awareness nowadays the consumption of products of organic agriculture is increasing. Organic farming means excluding the use of synthetic inputs, such as synthetic fertilizers and pesticides or genetically modified organisms, but there are only a few regulations regarding heavy metal concentrations in soils. Thus a not negligible uptake of metals from the soil where the apple trees are grown may occur. Furthermore inorganic copper compounds being traditional fertilizers for apple trees are not considered as synthetic fertilizers, thus they are still used in organic apple farming for soil or foliar application. Thus also apples produced by organic agriculture may contain toxic elements, such as cadmium, chromium, copper, and lead. The concentrations of these elements were determined in whole apples, as well as in the flesh and peel in order to estimate the possible risk for human health. Prior to analysis using ICP-AES samples underwent a microwave assisted digestion. The LODs obtained are below the recommended maximum levels in vegetables/fruits by WHO. In all samples no Cu, Cr, and Pb could be detected. Only Ca was found in the peel of about a third of samples investigated in concentrations between LOD and LOQ (1-3 μg/g).
基金Under the auspices of National Basic Research Program of China(No.2010CB951802,2005CB422207)Knowledge Innovation Program of Chinese Academy of Sciences(No.KZCX2-YW-219)National High Technology Research and Development Program of China(No.2009AA122104)
文摘The effects of acid deposition on pine forest ecosystems in Longli of Guizhou Province, southwestern China are studied using indoor experiments and model simulations. Indoor experiments are designed to explore the aluminum toxicity on pine seedlings, and the long-term soil acidification model (LTSAM) and a terrestrial biogeochemistry model (CENTURY) are used to simulate the influences of acid deposition on pine forest ecosystems. The indoor experiment results of aluminum toxicishow that aluminum ions in solution limit plant growth and acid deposition enhances this effect by facilitating the release of aluminum ions from the soil. Pine seedling bio- mass and root elongation decrease as the aluminum concentration increases. The results of model simulations show that the soil chemis- try varies significantly with different changes in acid deposition. When the acid deposition increases, the pH value in the soil solution decreases and the soil A13+ concentration increases. The increased acid deposition also has negative impacts on the forest ecosystem, i.e., decreases plant biomass, net primary productivity (NPP) and net C02 uptake. As a result, the soil organic carbon (SOC) decreases be- cause of the limited supply of decomposition material. Thus acid deposition need be reduced to help protect the forest ecosystems.
文摘AIM: To explore dendritic cells (DCs) multiple functions in immune modulation. METHODS: We used bone-marrow derived dendritic cells from BALB/c mice pulsed with pseudo particles from the hepatitis C virus to vaccinate naive BALB/c mice. Hepatitis C virus (HCV) pseudo particles consist of the genotype 1b derived envelope proteins E1 and E2, covering a non-HCV core structure. Thus, not a single epitope, but the whole "viral surface" induces immunogenicity. For vaccination, mature and activated DC were injected subcutaneously twice. RESULTS: Humoral and cellular immune responses measured by enzyme-linked immunosorbent assay and interferon-gamma enzyme-linked immunosorbent spot test showed antibody production as well as T-cellsdirected against HCV. Furthermore, T-cell responses confi rmed two highly immunogenic regions in E1 and E2 outside the hypervariable region 1. CONCLUSION: Our results indicate dendritic cells as a promising vaccination model for HCV infection that should be evaluated further.
文摘The deoiled Jatropha curcas meal by hexane extraction was detoxified phorbol esters by two different methods. These two methods were alkali in methanol and only ethanol washing. After both treatments, the PEs (phorbol esters) was decreased by 100%. The crude protein in detoxified meal of alkali in methanol washing was less amount than only ethanol washing. The result showed that treatment by only ethanol washing was a promising way to detoxify deoiled Jatropha curcas meal for animal feeds in industrial scale.
文摘Invasions by exotic insects and diseases are one of the most important threats to the stability and productivity of forest ecosystems around the world. Lymantria dispar (Lepidoptera: Lymantriidae) in Iran was observed for the first time in 1937 by Jalal Afshar in oak forests of the Guilan state region. Since its introduction, gypsy moth has spread to the northern, western and south-western regions of Iran. The largest outbreaks of gypsy moth occurred in the Talesh Oak forests of the Hyrcanian zone in 1975. The gypsy moth is a highly polyphagous folivore species that feeds on over 300 species of woody plants. Among its most preferred hosts are oaks and aspens. Some trees are resistant to the gypsy moth including honey locust, black locust, silver maple, green ash, dogwood, sycamore, horse chestnut, firs, and tulip trees. Defoliation during severe outbreaks alters the appearance of forest habitats and leads to tree mortality and changes in stand composition. This study was conducted during 2008 and 2009 in an effort to determine the primary host tree species that the gypsy moth infests in the Hyrcanian forests of lran. For that purpose all egg masses in the area were counted and conclusively significant differences in defoliation levels among tree species were found. The primary host tree species of gypsy moth in Iran was Persian ironweed, Parrotia persica.
文摘A two-step method was developed to quantitatively assess the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis (Li) Humber, on the green peach aphid, Myzus persicae (Sulzer) .Firstly, a standard time-dose-mortality relationship, established by modeling data from bioassay 1 at varying conidial dosages (0.4- 10.4 conidia/mm^2) of Z. anhuiensis F97028, was used to yield an estimate of expected mortality probability at a given dosage. Secondly, bioassay 2 was conducted by simultaneously exposing six ≤4-day-old nymphal colonies to a shower of Z. anhuiensis conidia at each of four dosages (resulting from exposures of 0.3 - 8.0 min) . Subsequently, the colonies were separately immersed in a 0.1% chlorothalonil solution for 0.5 min to disinfect all surviving conidia on the host integument from 1 - 12 h after exposure under temperature treatments of 15 and 20℃, respectively. The infection rate during a specific period from the end of the exposure to the immersion was then estimated as the ratio of the observed mortality over the expected mortality probability at a particular dosage. The results showed that the infection of M. persicae from Z. anhuiensis was highly rapid with little difference between aphid colonies maintained at 15 and 20℃ before being immersed in the fungicidal solution after exposure. The first 6-hour period after exposure was most crucial to successful infection of the fungus with the infection rate greatly depending on conidial dosages. It took ≤ 1 h to infect > 50% of the aphids at a dosage of > 1.5 conida/mm^2 and > 90% at > 50 conidia/mm^2 .
基金This research is supported by the Government of India, Ministry of Science and Technology, Mathematical Science Office, No. SR/S4/MS: 558/08 and supported partially by the National Natural Science Foundation of China (No. 11271314) and Plan for Scientific Innovation Talent of Henan Province (No. 144200510021).
文摘In this research paper, our main objective is to find out the meticulous role of activated dendritic cells (DCs) during the human immunodeficiency virus (HIV) infection process. DCs play a dual role by enhancing both HIV infection progression, as well as antiviral immune response. To explore the implications of these dual roles, we have formulated our mathematical model and analyzed the model by both analytical and numerical approaches. By using an impulsive differential equation, we have studied the effect of DC-based vaccination. Analytically we have determined the threshold value of drug dosage and dosing interval for optimum levels of injection. We have also investigated the effect of perfect adherence of drug dose on the immune cell count in extreme cases and observed that, systematic drug dose of the immune cells leads to its maximum level.
文摘A new technique for mass rearing Clostera anachoreta(C.) was developed through a large numberof experiments, so the problem of mass produce of CaGV was solved. The process of CaGV was discussedin detail.