Mir204 and Mir211 suppress synovial inflammation and proliferation in rheumatoid arthritis by targeting Ssrp1.

Rheumatoid arthritis (RA) is a chronic inflammatory joint disease characterized by synovial hyperplasia. Mir204 and Mir211 are homologous miRNAs with the same gene targeting spectrum. It is known that Mir204/211 play an important role in protecting osteoarthritis development; however, the roles of Mir204/211 in RA disease have not been determined. In the present study, we investigated the effects and molecular mechanisms of Mir204/211 on synovial inflammation and hyperproliferation in RA. The effects of Mir204/211 on the inflammation and abnormal proliferation in primary fibroblast-like synoviocytes (FLSs) were examined by Mir204/211 gain-of-function and loss-of-function approaches in vitro and in vivo. We identified the structure-specific recognition protein 1 (Ssrp1) as a downstream target gene of Mir204/211 based on the bioinformatics analysis. We overexpressed Ssrp1and Mir204/211 in FLS to determine the relationship between Ssrp1 and Mir204/211 and their effects on synovial hyperplasia. We created a collagen-induced arthritis (CIA) model in wild-type as well as Mir204/211 double knockout (dKO) mice to induce RA phenotype and administered adeno-associated virus (AAV)-mediated Ssrp1-shRNA (AAV-shSsrp1) by intra-articular injection into Mir204/211 dKO mice. We found that Mir204/211 attenuated excessive cell proliferation and synovial inflammation in RA. Ssrp1 was the downstream target gene of Mir204/211. Mir204/211 affected synovial proliferation and decelerated RA progression by targeting Ssrp1. CIA mice with Mir204/211 deficiency displayed enhanced synovial hyperplasia and inflammation. RA phenotypes observed in Mir204/211 deficient mice were significantly ameliorated by intra-articular delivery of AAV-shSsrp1, confirming the involvement of Mir204/211-Ssrp1signaling during RA development. In this study, we demonstrated that Mir204/211 antagonize synovial hyperplasia and inflammation in RA by regulation of Ssrp1. Mir204/211 may serve as novel agents to treat RA disease.

Pharmacology and molecular docking study of cartilage protection of Chinese herbal medicine Fufang Shatai Heji (STHJ) by inhibiting the expression of MMPs in collagen-induced arthritis mice.

BACKGROUND: This study aims to explore whether Fufang Shatai Heji (STHJ), as a mixture collected by a decoction of a variety of Chinese herbal medicines for immune system diseases, can improve the cartilage destruction of rheumatoid arthritis (RA). METHODS: The therapeutic effects of STHJ were studied using collagen induced arthritis (CIA) mice. The improvement effect of STHJ on synovitis and cartilage damage caused by arthritis was studied by joint pathological analysis. The inhibitory effect of STHJ on related degradation enzymes in cartilage was studied by immunohistochemistry and real-time polymerase chain reaction (PCR). The specific targets of STHJ were predicted by molecular docking. RESULTS: After successfully inducing CIA, the paws of the mice showed significant swelling, and athological analysis of the ankle and knee joints also showed significant cartilage destruction and synovial hyperplasia. However, synovial hyperplasia and cartilage destruction were markedly alleviated after administration of STHJ. And after STHJ treatment, the expression of ADAMTS-4, ADAMTS-5, MMP-9 and MMP-13, in the cartilage layer of CIA mice was significantly inhibited. Through molecular docking assays, we proved that acteoside in STHJ could directly bind to the Glu111, Phe110 residues in MMP-9 and glycyrrhizic acid in STHJ bind to the Glu382, Asn433 residues in MMP-13. CONCLUSIONS: Our results suggested that STHJ may alleviate synovial hyperplasia and cartilage destruction in CIA mice and protect cartilage by inhibiting the expression of MMP-9 and other enzymes.

Dexamethasone-loaded thermo-sensitive hydrogel attenuates osteoarthritis by protecting cartilage and providing effective pain relief.

BACKGROUND: We utilized the destabilization of medial meniscus (DMM)-induced mice to illustrate the osteoarthritis (OA) suppressing and pain-relieving effects of a novel prolonged-release intra-articular (IA)-dexamethasone-loaded thermo-sensitive hydrogel (DLTH). METHODS: The effects of temperature and pH on DLTH formation and in vitro DLTH release profile were assessed. C57BL/6J mice were randomly divided into three groups: Ctrl group, Model group and DLTH group. The DLTH group received joint injections of 10 µL DLTH (1 mg/kg) into the right knee once a week from week 2 to week 11. We performed micro-computed tomography (Micro-CT) and histological analyses of safranin O-fast green, hematoxylin and eosin, and tartrate-resistant acid phosphatase in knee joints. We also carried out immunohistochemical (IHC) staining for matrix metalloproteinase-9 (MMP-9), MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) in cartilage and Ki-67 in synovia. Pain behavioral testing was carried out in all mice. The serum content of prostaglandin E2 (PGE2) and real-time polymerase chain reaction (PCR) of inflammatory cytokines and pain-related factors in dorsal root ganglia (DRGs) were evaluated. RESULTS: It took 20 minutes to form DLTH at pH 7.0 and 37 °C. The cumulative release profiles of dexamethasone (Dex) from DLTH at 37 °C revealed a rapid release in the first 24 h and a sustained slow release for 7 days. In vivo study illustrated that DLTH attenuated OA bone destruction and ameliorated synovitis and progression of OA in DMM-induced mice. The chondroprotective effects of DLTH were mediated by decreased expressions of MMP-9, MMP-13, and ADAMTS-5. The results showed that IA-DLTH exerted pain-relieving effects in OA mice. Upregulation of nociceptive response time (NRT) and downregulations of serum PGE2, inflammatory factors, and pain-related mediators in DRGs of mice in the DLTH group were recorded. CONCLUSIONS: Data presented in this study elucidated that DLTH exhibited a long and lasting Dex release and it is a potential sustainable drug delivery system (DDS) to treat OA locally. IA-DLTH injection exerted chondroprotective and pain-relieving effects in DMM-induced arthritis. The involvement of MMP-9, MMP-13, ADAMTS-5, and inflammatory and pain-related factors, may account for the suppression of OA progression and pain.

Establishment and Evaluation of a Novel High-Efficiency Model of Graded Traumatic Brain Injury in Mice.

BACKGROUND: Although previous studies have made significant contributions to establishing animal traumatic brain injury (TBI) models for simulation of human TBI, the accuracy, controllability, and modeling efficiency of animal TBI models need to be further improved. This study established a novel high-efficiency graded mouse TBI model induced by shock wave. METHODS: A total of 125 mice were randomly divided into sham, 0.7 mm, 0.6 mm, and 0.5 mm groups according to the depth of the cross groove of the aluminum sheets. The stability and repeatability of apparatus were evaluated, and the integrity of the blood-brain barrier, cerebral edema, neuropathologic immunohistochemistry, apoptosis-related protein, and behavioral tests of neurologic function were used to validate this new model. RESULTS: The results showed that 4 mice were injured simultaneously in 1 experiment. They received the same intensity of shock waves. Moreover, the mortality rates caused by 3 different aluminum sheets were consistent with the mortality rates of mild TBI, moderate TBI, and severe TBI. Compared with the sham group, mice in different injured groups significantly increased brain water content, blood-brain barrier permeability, and neuronal apoptosis. And the mice in all injured groups showed poor motor ability, balancing, spatial learning, and memory abilities. CONCLUSIONS: The novel TBI apparatus has advantages in its small size, simple operation, high repeatability, high efficiency, and graded severity. Our TBI apparatus provides a novel tool to investigate the neuropathologic changes and underlying mechanisms of TBI with various levels of severities.

Dexamethasone-Loaded Thermosensitive Hydrogel Suppresses Inflammation and Pain in Collagen-Induced Arthritis Rats.

PURPOSE: To overcome negative adverse effects and improve therapeutic index of dexamethasone (Dex) in rheumatoid arthritis (RA), we developed a novel sustained release formulation-intra-articular injectable dexamethasone-loaded thermosensitive hydrogel (DLTH) with chitosan-glycerin-borax as carrier for the remission of inflammation and pain. The focus of this article is to explore both anti-inflammatory and pain-relieving effects of DLTH joint injection in bovine type-II collagen-induced arthritis (CIA) rats. METHODS: Wistar rats were randomized into three groups, including the normal group (n=6), the model group (n=6) and the DLTH group (n=10). Joint injection of DLTH (1mg/kg Dex per rat) was injected on day 12 in the DLTH group twice a week for three weeks. Clinical signs of body weight, paw swelling and arthritis scores, histologic analysis, hind paw mechanical withdrawal threshold (MWT), plantar pressure pain threshold (PPT) were taken into consideration. Serum contents of IL-17A, prostaglandin E2 (PGE2), prostacyclin 2 (PGI2) and prostaglandin D2 (PGD2), real-time polymerase chain reaction (PCR) analysis of inflammatory factors and pain-related mediators in synovium and dorsal root ganglia (DRG), Western blotting of NF-κB in synovium were all evaluated. RESULTS: Paw swelling, arthritis scores and joint inflammation destruction were all attenuated in the DLTH-treated group. Results showed that DLTH not only down-regulated serum IL-17A, but also mRNA levels of inflammatory factors and NGF, and key proteins contents of the NF-κB pathway in synovium. Increases of MWT and PPT in DLTH-treated rats elucidated pain-reducing effects of DLTH. Elevated serum PGD2 levels and declines of serum PGE2 and PGI2, and inflammatory and pain-related genes in DRGs in the DLTH group were also recorded. CONCLUSION: These data elucidated that DLTH joint injection impeded synovial inflammation processes through down-regulating transcription activity of NF-κB pathway, and intra-articular DLTH may aid in the regulation of RA pain through regulating inflammation and pain conduction process.

Correction to: Simulated microgravity significantly altered metabolism in epidermal stem cells.

The affiliation given for Yan Cui in this article is not correct. The following is the correction affiliation.

[Collagen Peptides Improve Lymphocyte Distribution in Peripheral Blood and T Lymphocyte Proliferation in Spleen of Mice under the Condition of Simulated Weightlessness].

OBJECTIVE: To investigate whether collagen peptides can improve the immune functions of mice under the condition of simulated weightlessness. METHODS: Mouse tail-suspension model was used to simulate the effects of weightlessness. Tail-suspended mice were intraperitoneally injected with 600 mg collagen peptides per kilogram body weight once a day for 10 days. Then, the mice were killed, and white blood cells were counted and classified. Lymphocyte subsets and T lymphocyte proliferations in spleens were analyzed. RESULTS: Compared with normal control group, total and differential count of leukocytes, lymphocytes, T cells，CD4+ and CD8+ T cells, B cells and NK cells, and splenic T lymphocyte proliferation all decreased in the weightlessness simulated mice (P＜0.05). Except for NK cells, the above-mentioned parameters were increased after administration of collagen peptides, and some of the parameters were recovered to the levels of normal control mice (P＜0.05). CONCLUSION: Collagen peptides can effectively improve peripheral blood lymphocyte distributions and T lymphocyte proliferations of mice under the condition of simulated weightlessness. This study nay provid the experimental basis for improvement of immune functions of astronauts.

DMY protects the knee joints of rats with collagen-induced arthritis by inhibition of NF-κB signaling and osteoclastic bone resorption.

Collagen-induced arthritis (CIA) is a widely used animal model for studying rheumatoid arthritis (RA), which manifests serious joint dysfunction, progressive bone erosion and articular cartilage destruction. Considering that joint damage in RA is caused by systemic inflammation and dihydromyricetin (DMY), the main flavonoid of Ampelopsis Michx, possesses anti-inflammatory properties, in the present study we have investigated the potential capability of DMY to inhibit inflammation-mediated joint damage and explore the underlying mechanisms. A rat model of RA induced by CIA was administered with DMY for 5 weeks. Prior to histological analysis, the knee joints were scanned by microcomputed tomography (µCT) to detect bone damage. Articular cartilage destruction was assessed by Alcian blue and Toluidine blue staining and the pathological alteration of osteoblasts and osteoclasts in joints was evaluated by hematoxylin-eosin (H&E) and tartrate-resistant acid phosphatase (TRAP) staining, respectively. The effects of DMY on osteoblast differentiation and osteoclast formation in vitro were investigated. Consistent with the in vivo results, DMY had no significant effect on osteoblast differentiation but an inhibitory effect on osteoclast formation. Furthermore, we determined that the mechanism of the DMY-suppressed osteoclast formation was blocking the phosphorylation of I-κB kinase (IKK) so as to hinder the activation of nuclear factor-κB (NF-κB). Collectively, DMY could ameliorate knee joint damage, especially in articular cartilage, which is the weight-bearing region, by inhibiting osteoclast formation through NF-κB signaling.

Simulated microgravity significantly altered metabolism in epidermal stem cells.

Simulated microgravity can significantly affect various cell types and multiple systems of the human body, such as cardiovascular system, skeletal muscle system, and immune system, and is known to cause anemia and loss of electrolyte and fluids. Epidermal stem cells (EpSCs) were cultured in a rotary cell culture system (RCCS) bioreactor to simulate microgravity. The metabolites of EpSCs were identified by liquid chromatography-mass spectrometry (LC-MS). Compared with normal gravity (NG) group, a total of 57 different metabolites of EpSCs were identified (P < 0.05, VIP > 1), including lipids and lipid-like molecules (51 molecules), amino acids (5 molecules), nucleosides, nucleotides, and analogues (1 molecule). According to the partial least squares discriminant analysis (PLS-DA) score plot, a VIP > 1 and P < 0.05 were obtained for the 57 different metabolites, of which 23 molecules were significantly downregulated and 34 were significantly upregulated in simulated microgravity (SMG) group. These results showed that SMG has a significant impact on different pathways, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated that multiple pathways were involved, mainly the amino acid metabolism pathway, lipid metabolism pathway, membrane transport pathway, and cell growth and death pathways. Thus, the metabolic profile of EpSCs was changed under SMG. Exploring the metabolic profile of EpSCs would be helpful to further understand the growth characteristics of EpSCs under SMG, which will provide a new approach to explore the metabolomics mechanism of stress injury and repair trauma under SMG.

The Traditional Chinese Medicine Fufang Shatai Heji (STHJ) Enhances Immune Function in Cyclophosphamide-Treated Mice.

Fufang Shatai Heji (STHJ) is a mixture of traditional Chinese medicines, such as Radix Adenophorae, Radix Pseudostellariae, and Radix Astragali. STHJ is commonly used to treat diseases caused by low immune function, for example, Sjögren's syndrome (SS). The primary objective of this study was to assess the immunopotentiating effect of STHJ using an immunosuppressive mouse model receiving cyclophosphamide (CTX). Following CTX treatment, STHJ was administered by oral gavage for 30 consecutive days. The percentage of specific lymphocyte subpopulations in the spleen was measured by flow cytometry. Levels of inflammatory factors in serum were detected by enzyme-linked immunosorbent assays (ELISAs). The administration of STHJ significantly elevated thymus and spleen indices, increased B cell and natural killer (NK) cell activities, and decreased CD8+ T, CD8+CD122+ T, NKT, and γδT cell activities in the CTX-treated mice. In addition, STHJ upregulated the expression of interleukin- (IL-) 2, IL-6, and tumor necrosis factor-α (TNF-α) and downregulated IL-10 expression in CTX-treated mice. In conclusion, STHJ effectively remitted CTX-induced immunosuppression by modulating the balance of lymphocyte subsets and cytokines. Our results suggest STHJ treatment could be used as an effective therapeutic approach to improve immune function in patients with low immunity.

Metformin inhibits inflammation and bone destruction in collagen-induced arthritis in rats.

BACKGROUND: Metformin (MF) is a widely used biguanide oral hypoglycemic agent, which has obvious anti-inflammatory and immunomodulatory effects. However, the mechanism of MF on rheumatoid arthritis (RA) remains uncertain. In this study, we investigated the therapeutic effects of MF on collagen-induced arthritis (CIA). METHODS: CIA was induced in rats by intradermal injection of a mixture of bovine type II collagen and incomplete Freund's adjuvant (IFA) on day 0 and day 7 through the base of the tail. Intraperitoneal injection of MF (100 mg/kg) was given every 3 days, from day 14 for 3 weeks. The effects of MF on arthritis-induced systemic inflammation and synovitis were studied by pathological analysis of the knee joint and serological examination of peripheral blood in CIA rats. The bone protection effect of MF was studied by microscopic computed tomography (micro-CT) and histological analysis of the knee joint. The effects of MF on chondrocytes in CIA rats were studied by detecting the relevant pro-apoptotic mediators in the chondrocytes. RESULTS: After administration of MF in CIA rats, systemic inflammation and synovitis caused by arthritis were significantly suppressed. Histomorphometry and micro-CT analysis of the knee joint revealed that MF can protect bone by inhibiting the changes of trabecular bone in CIA rats. Histological analysis of the knee joint found that MF can inhibit osteoclast formation and degradation of the cartilage layer matrix. Detection of the relevant pro-apoptotic mediators in chondrocytes revealed that MF can significantly inhibit the apoptosis of chondrocytes in CIA rats. CONCLUSIONS: Our study showed that MF can inhibit systemic inflammation and synovitis and plays a role in bone protection by inhibiting cartilage layer matrix degradation, osteoclast formation, and chondrocyte apoptosis.

Recombinant adenovirus of SEA and CD80 genes driven by MMRE and mouse TERT promoter induce effective antitumor immune responses against different types of tumor cells in vitro and in vivo.

Staphylococcus enterotoxin A (SEA) is a powerful immunostimulant and can stimulate T cells bearing certain T-cell receptor ß-chain variable regions when bound to major histocompatibility complex II molecules. SEA is widely used in research of antitumor therapy. The low affinity T-cell receptor (TCR) interaction with SEA in the absence of MHC class II antigens is sufficient for the induction of cytotoxicity but requires additional CD28/B7 signaling to result in proliferation of resting T cells. In this study, we constructed recombinant adenovirus (named as Ad-MMRE-mTERT-BIS) carrying membrane-expressing SEA (named as SEAtm) and CD80 driven by Myc-Max response elements (MMRE) and mouse telomerase reverse transcriptase (mTERT) promoter to reduce toxicity and to improve safety and efficiency. We demonstrated that Ad-MMRE-mTERT-BIS could make SEAtm and CD80 to co-express highly on the surface of Hepa1-6 and B16 cells, at low level on the surface of CT26 cells, but not in NIH3T3. Hepa1-6 and B16 cells infected by the recombinant adenovirus induced proliferation of CD4+ and CD8+ T cells and increased cytokine [interleukin (IL)-2, tumor necrosis factor (TNF)-α, interferon (IFN)-γ] production in vitro. Intratumoral injection of Ad-MMRE-mTERT-BIS in hepatoma and melanoma mouse models induced tumor-specific cytotoxic T cells in the spleen. Moreover, hepatoma and melanoma xenografts were suppressed by treatment with Ad-MMRE-mTERT-BIS and the survival time of treated mice was prolonged. These findings suggest that recombinant adenovirus of SEA and CD80 genes driven by mTERT promoter could induce effective antitumor immune responses against different kinds of tumor cells in vitro and in vivo.

[Effects of sand burial on fluxes of greenhouse gases from the soil covered by biocrust in an arid desert region.] / 沙埋对干旱沙区生物结皮覆盖土壤温室气体通量的影响.

Based on the measurements of the fluxes of CO2, CH4 and N2O from the soil covered by two types of biocrusts dominated separately by moss and algae-lichen, followed by 0 (control), 1 (shallow) and 10 (deep) mm depths of sand burial treatments, we studied the effects of sand burial on greenhouse gases fluxes and their relationships with soil temperature and moisture at Shapotou, southeastern edge of the Tengger Desert. The results showed that sand burial had significantly positive effects on CO2 emission fluxes and CH4 uptake fluxes of the soil covered by the two types of biocrusts, but imposed differential effects on N2O fluxes depending on the type of biocrust and the depth of burial. Deep burial (10 mm) dramatically increased the N2O uptake fluxes of the soil co-vered by the two types of biocrusts, while shallow burial (1 mm) decreased the N2O uptake flux of the soil co-vered by moss crust only and had no significant effects on N2O uptake flux of the soil covered by algae-lichen crust. In addition, CO2 fluxes of the two biocrusts were closely related to the soil temperature and soil moisture, thereby increasing with the raised soil surface temperature and soil moisture caused by sand burial. However, the relationships of burial-induced changes of soil temperature and moisture with the changes in the other two greenhouse gases fluxes were not evident, indicating that the variations of soil temperature and moisture caused by sand burial were not the key factors affecting the fluxes of CH4 and N2O of the soil covered by the two types of biocrusts.

[Nitrification of biological soil crusts and soil system during drought process and its response to temperature and moisture: A case study in the Shapotou region, Northwest China].

Two types of soil covered by biological soil crusts (BSCs) , i.e. moss and algae, and moving sand in the natural vegetation area at the southeast fringe of the Tengger Desert were collected intactly. They were incubated continuously for 20 days under two different temperatures (15 degrees C and 25 degrees C) and moistures (10% and 25%) condition in the laboratory, and soil NO3(-)-N contents were measured after 1, 2, 5, 8, 12, 20 days of incubation and net nitrification rate was evaluated during dehydration. The results showed that NO3(-)-N content of the moss-covered soil (2.29 mg x kg(-1)) was higher than that of the algae-covered soil (1.84 mg x kg(-1)) and sand (1.59 mg x kg(-1)). Net nitrification rate of the three soil types ranged from -3.47 to 2.97 mg x kg(-1) x d(-1). For the moss-covered soil and algae-covered soil at 10% and 25% moisture levels, the net nitrification rates at 15 degrees C were 75.1%, 0.7% and 99.1%, 21.3% higher than those at 25 degrees C, respectively. Also, the net nitrification rates at 15 degrees C and 10% moisture levels were 193.4% and 107.3% higher than those at 25 degrees C and 25% moisture levels, respectively. The results suggested that regardless of soil moisture increasing or decreasing under the global warming senior, the net nitrification rate of BSCs-soil system in the desert would probably be limited to some extent during drought process.

[Greenhouse gases fluxes of biological soil crusts and soil ecosystem in the artificial sand-fixing vegetation region in Shapotou area].

Uncertainties still existed for evaluating greenhouse gases fluxes (GHGs), including carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) at the regional scale for desert ecosystem because available GHGs data about biological soil crusts (BSCs) was very scarce. In 2011 and 2012, soil ecosystem covered by various types of BSCs and BSCs at different succession stages in an artificial sand-fixing vegetation region established in various periods at southeast of the Shapotou area in Tengger Desert was selected to measure fluxes of CO2, CH4 and N2O using static chamber and gas chromatography. The results showed that curst type, recovery time and their interactions with sampling date significantly affected CO2 flux. Recovery time and interaction of crust type and sampling date significantly affected CH4 flux. Sampling date significantly affected the fluxes of CO2, CH4 and N2O. The mean annual flux of CO2 for moss crust (105.1 mg x m(-2) x h(-1)) was significantly higher than that of algae crust (37.7 mg x m(-2) x h(-1)) at the same succession stage. Annual mean CH4 and N2O consumption was 19.9 and 3.4 microg x m(-2) x h(-1), respectively. Mean annual consumption of CH4 and N2O for algae crust was slightly higher than that of moss crust, however, significant difference was not found. Ecosystem respiration (Re) of desert soil covered by BSCs increased with the recovery process of desert ecosystem, in contrast, consumption of CH4 and N2O decreased. Re of moss crust was more sensitive to temperature and moisture variation than algae crust and Re sensitivity of temperature and moisture gradually increased with the development and succession of BSCs. Both soil temperature and moisture were not the main factor to determine CH4 and N2O fluxes of BSCs-soil in desert ecosystem.

[Changes of anti-Streptococcus pneumoniae status in rats with simulated weightlessness].

OBJECTIVE: To study the changes of anti-Streptococcus pneumonia (SP) status in rats with simulated weightlessness, and therefore to provide theoretical basis for the aerospace medicine. METHODS: Thirty-two healthy male Wistar rats were randomly allocated into 4 groups: group A, the tail-suspension and SP group; group B, the tail-suspension without SP group; group C, the unsuspended but SP group; group D, the unsuspended and no SP group, with 8 rats in each. The tail-suspension method, i.e. about 30° head-down tilt, was used for the model of simulated microgravity. On day 4, 0.4 ml of SP suspension [ATCC6303, serotype III(ATCC, bacteria concentration about 9.0×108 CFU/ml)] was instilled by tracheal intubation. Sterile saline was used for the control group. The experiment was ended after 7 days of tail-suspension. Lung pathology, blood test and C-reactive protein level were studied, and the CD(4)(+)/CD(8)(+) ratios were measured by flow cytometry. RESULTS: The lung pathological changes were much more severe in Group A as compared to those in Group B, C and D. The total number of WBC showed no significant difference among groups (F = 1.57, P = 0.22). But the neutrophil number was higher in Group A [(2.4 ± 0.53)×109/L], B [(2.0 ± 0.31)×109/L] and C [(1.7 ± 0.40)×109/L] as compared to Group D [(1.2 ± 0.15)×109/L], u = 0.0001, P = 0.001; u = 1.0, P = 0.001; u = 8.5, P = 0.013, respectively. The percentage of neutrophils showed a similar difference. The total number of lymphocytes showed no significant difference among groups (F = 0.720, P = 0.548). CRP levels in the SP infection groups were significantly higher than those in the uninfected groups. The ratio of CD(4)(+)/CD(8)(+) showed no difference among groups (F = 1.225, P = 0.319). Weight loss after the experiment was most severe in Group A (F = 122.067, P < 0.001). CONCLUSIONS: In rats with simulated weightlessness, the anti-infective ability to Streptococcus pneumoniae was reduced, and the inflammatory response was significantly increased, but the anti-infective immunity was compromised.

[Construction of recombinant adenovirus of SEA and CD80 genes co-expression regulated by mouse TERT promoter and identification of its expression in hepatoma cells].

AIM: To construct recombinant co-expression adenovirus vector of SEA and CD80 genes regulated by mouse TERT(telomerase reverse transcriptase, TERT) promoter and to observe the expression of SEA and CD80 in the Hepa1-6 cells mediated by it. METHODS: Using AdEasy adenovirus system, the core promoter region of mTERT was subcloned to shuttle plasmid pShuttle2 and Myc-Max response element was inserted upstream of it to regulate the expression of SEA and CD80. The recombinant co-expression adenovirus vector of SEA and CD80 genes was constructed and named as Ad-MMRE-mTERT-BIS. Hepatoma cell line Hepa1-6 and fibroblast cell line NIH3T3 were infected by recombinant adenovirus at MOI(multiplicity of infection)of 100, the expression of SEA and CD80 on the surface of cells was detected by indirect immunofluorescent staining. RESULTS: SEA and CD80 was specifically co-expressed on the surface of infected Hepa1-6 cells but not on NIH3T3 cells. CONCLUSION: The recombinant co-expression adenovirus vector of SEA and CD80 gene regulated by mTERT promoter was sucessfully constructed and make targeting-expression of SEA and CD80 on the surface of hepatoma cells, which lays the foundation for further research on application of SEA and CD80 in targeted genetherapy for hepatoma.

Microarray analysis of altered gene expression in diallyl trisulfide-treated HepG2 cells.

Diallyl trisulfide (DT) is a natural compound derived from garlic. Despite its reported lipid-lowering effects, the mechanisms of its actions are not clear yet. To further understand the molecular mechanisms of actions of this compound, microarray technology was used in the present study to investigate the lipid-lowering effects of DT on HepG2 cells. To optimize the concentration of DT treatment on HepG2 cells, a series of concentration of DT were incubated with HepG2 cells for 24 h. The data indicted that the concentrations of DT in the range from 20 to 50 microM were effective in lowering cellular total triglyceride and cholesterol, with no significant cytotoxicity. Using oligonucleotide microarrays and RT-PCR technology, the genes that were differentially regulated by DT were identified. The results showed that peroxisome proliferator activated receptor alpha (PPAR-alpha) and hepatocyte nuclear factor 4alpha (HNF-4alpha) mRNA were up-regulated, and CYP7A1 mRNA was down-regulated following DT treatment, suggesting that the lipid-lowering effects of DT may be at least in part mediated through the regulation of PPAR-alpha dependent pathways.