IL-37b gene transfer enhances the therapeutic efficacy of mesenchumal stromal cells in DSS-induced colitis mice.

AIM: To investigate whether the transfer of the IL-37b gene, a newly identified inhibitor of both innate and adaptive immunity, could improve the therapeutic efficacy of mesenchumal stromal cells (MSCs) in inflammatory bowel disease (IBD). METHODS: The expression of IL-37 in biopsied specimens of the patients with active ulcerative colitis (UC) was detected using RT-PCR and immunohistochemistry. Mice were treated with 3% dextran sulfate sodium (DSS) for 8 days to induce colitis. Before DSS treatment, the mice were injected with MSCs, MSC-eGFP or MSC-IL37b. Their body weight was measured each day, and the colons and spleens were harvested on d 10 for pathological and biochemical analyses. RESULTS: In biopsied specimens of the patients with active UC, the expression of IL-37 was dramatically elevated in inflamed mucosa, mainly in epithelial cells and infiltrating immune cells. Compared to MSC-eGFP or MSCs, MSC-IL37b administration significantly attenuated the body weight and colon length reduction, and decreased the histological score in DSS-induced colitis mice. Furthermore, MSC-IL37b administration increased the percentage of myeloid-derived suppressor cells (MDSCs) among total splenic mononuclear cells as well as the percentage of regulatory T cells (Tregs) among splenic CD4+ T cells in the mice. Moreover, MSC-IL37b administration increased the IL-2+ cells and decreased the IFN-γ+ cells among splenic CD4+ T cells. CONCLUSION: IL-37 is involved in the pathophysiology of UC. IL-37b gene transfer enhances the therapeutic efficacy of MSCs in DSS-induced colitis mice by inducing Tregs and MDSCs and regulating cytokine production.

Transfer of the IL-37b gene elicits anti-tumor responses in mice bearing 4T1 breast cancer.

AIM: IL-37b has shown anti-cancer activities in addition to its anti-inflammatory properties. In this study, we investigated the effects of IL-37b on breast carcinoma growth in mice and to determine the involvement of T cell activation in the effects. METHODS: IL-37b gene was transferred into mouse breast carcinoma cell line 4T1 (4T1-IL37b cells), the expression of secretory IL-37b by the cells was detected, and the effects of IL-37b expression on the cell proliferation in vitro was evaluated. After injection of 4T1 cells or 4T1-IL37b cells into immunocompetent BALB/c mice, immunodeficient BALB/c nude mice and NOD-SCID mice, the tumor growth and survival rate were measured. The proliferation of T cells in vitro was also detected. RESULTS: IL-37b was detected in the supernatants of 4T1-IL37b cells with a concentration of 12.02 ± 0.875 ng/mL. IL-37b expression did not affect 4T1 cell proliferation in vitro. BALB/c mice inoculated with 4T1-IL37b cells showed significant retardation of tumor growth. BALB/c mice inoculated with both 4T1 cells and mitomycin C-treated 4T1-IL37b cells also showed significant retardation of tumor growth. But the anti-cancer activity of IL-37b was abrogated in BALB/c nude mice and NOD-SCID mice inoculated with 4T1-IL37b cells. Recombinant IL-37b slightly promoted CD4(+) T cell proliferation without affecting CD8(+) T cell proliferation. CONCLUSION: IL-37b exerts anti-4T1 breast carcinoma effects in vivo by modulating the tumor microenvironment and influencing T cell activation.

A dsRNA-binding protein MdDRB1 associated with miRNA biogenesis modifies adventitious rooting and tree architecture in apple.

Although numerous miRNAs have been already isolated from fruit trees, knowledge about miRNA biogenesis is largely unknown in fruit trees. Double-strand RNA-binding (DRB) protein plays an important role in miRNA processing and maturation; however, its role in the regulation of economically important traits is not clear yet in fruit trees. EST blast and RACE amplification were performed to isolate apple MdDRB1 gene. Following expression analysis, RNA binding and protein interaction assays, MdDRB1 was transformed into apple callus and in vitro tissue cultures to characterize the functions of MdDRB1 in miRNA biogenesis, adventitious rooting, leaf development and tree growth habit. MdDRB1 contained two highly conserved DRB domains. Its transcripts existed in all tissues tested and are induced by hormones. It bound to double-strand RNAs and interacted with AtDCL1 (Dicer-Like 1) and MdDCL1. Chip assay indicated its role in miRNA biogenesis. Transgenic analysis showed that MdDRB1 controls adventitious rooting, leaf curvature and tree architecture by modulating the accumulation of miRNAs and the transcript levels of miRNA target genes. Our results demonstrated that MdDRB1 functions in the miRNA biogenesis in a conserved way and that it is a master regulator in the formation of economically important traits in fruit trees.

Obesity and depressive symptoms in the elderly: a survey in the rural area of Chizhou, Anhui province.

OBJECTIVE: The aim of the present study is to examine the relationship between obesity and depressive symptoms and to test the "Jolly Fat" hypothesis among older Chinese. METHODS: A total of 736 rural Chinese aged 60 years and older participated in this cross-sectional study. Body mass index (BMI = kg/m(2) ) was calculated from the subjects' measured weight (kg) and height (meter). Depressive symptoms were assessed using the 30-item Geriatric Depression Scale (GDS-30), with a cut-off point of 11. RESULTS: Among 736 total participants, the prevalence of depressive symptoms was 24.1% in men and 27.9% in women. A trend about depressive symptoms decreased with increasing BMI was found in men (χ(2) trend = 5.74, df = 1, p = 0.01). A weak inverse linear trend between obesity and depressive symptoms was observed among subjects. In men, obese group was less likely to suffer from depressive symptoms compared with normal weight group before or after adjustment for confounders, with odds ratios of 0.32 (95% confidence interval (CI): 0.12-0.85) and 0.28 (95% CI: 0.09-0.85), respectively. However, the association between BMI and depressive symptoms in women showed no significant differences. CONCLUSIONS: Our results supported the "Jolly Fat" hypothesis only in rural older Chinese men, but not in women. Gender differences existed in the relationship between obesity and depressive symptoms.

Mesenchymal stem cells support proliferation and terminal differentiation of B cells.

BACKGROUND: Mesenchymal stem cells (MSC) play important roles in modulating the activities of T lymphocytes, dendritic cells and natural killer cells. These immunoregulatory properties of MSC suggest their therapeutic potential in autoimmune diseases. However, the effects of MSC on B cells are still poorly understood. The present study was designed to investigate the interaction between MSC and B cells both in vitro and in vivo, and to determine the possible mechanism of action. DESIGN AND METHOD: The effect of human umbilical cord mesenchymal stem cells (UC-MSC) on proliferation and differentiation of B-cells were characterized in vitro, and we also tested the immunoregulatory properties of mouse bone marrow MSC (BM-MSC) on T cell dependent and independent antibody production in vivo in mice. RESULTS: Treatment with human UC-MSC resulted in an increase of proliferation, differentiation of B cells into plasma cells and production of antibodies in vitro. Mouse BM-MSC significantly enhanced T cell dependent and independent antibodies production in vivo in mice. PGE2 partially mediated the immunosuppressive activity of human UC-MSC but IL-6 did not regulate this activity. CONCLUSION: MSC promote proliferation and differentiation of B cells in vitro and in vivo partially through PGE2 but not IL-6.

The cold-induced basic helix-loop-helix transcription factor gene MdCIbHLH1 encodes an ICE-like protein in apple.

BACKGROUND: Plant growth is greatly affected by low temperatures, and the expression of a number of genes is induced by cold stress. Although many genes in the cold signaling pathway have been identified in Arabidopsis, little is known about the transcription factors involved in the cold stress response in apple. RESULTS: Here, we show that the apple bHLH (basic helix-loop-helix) gene MdCIbHLH1 (Cold-Induced bHLH1), which encodes an ICE-like protein, was noticeably induced in response to cold stress. The MdCIbHLH1 protein specifically bound to the MYC recognition sequences in the AtCBF3 promoter, and MdCIbHLH1 overexpression enhanced cold tolerance in transgenic Arabidopsis. In addition, the MdCIbHLH1 protein bound to the promoters of MdCBF2 and favorably contributed to cold tolerance in transgenic apple plants by upregulating the expression of MdCBF2 through the CBF (C-repeat-binding factor) pathway. Our findings indicate that MdCIbHLH1 functions in stress tolerance in different species. For example, ectopic MdCIbHLH1 expression conferred enhanced chilling tolerance in transgenic tobacco. Finally, we observed that cold induces the degradation of the MdCIbHLH1 protein in apple and that this degradation was potentially mediated by ubiquitination and sumoylation. CONCLUSIONS: Based on these findings, MdCIbHLH1 encodes a transcription factor that is important for the cold tolerance response in apple.

Immunization of mice by hollow mesoporous silica nanoparticles as carriers of porcine circovirus type 2 ORF2 protein.

BACKGROUND: Porcine circovirus type 2 (PCV2) is a primary etiological agent of post-weaning multi-systemic wasting syndrome (PMWS), which is a disease of increasing importance to the pig industry worldwide. Hollow mesoporous silica nanoparticles (HMSNs) have gained increasing interest for use in vaccines. METHODS: To study the potential of HMSNs for use as a protein delivery system or vaccine carriers. HMSNs were synthesized by a sol-gel/emulsion(oil-in-water/ethanol) method, purified PCV2 GST-ORF2-E protein was loaded into HMSNs, and the resulting HMSN/protein mixture was injected into mice. The uptake and release profiles of protein by HMSNs in vitro were investigated. PCV2 GST-ORF2-E specific antibodies and secretion of IFN-γ were detected by enzyme-linked immunosorbent assays, spleen lymphocyte proliferation was measured by the MTS method, and the percentage of CD4+ and CD8+ were determined by flow cytometry. RESULTS: HMSNs were found to yield better binding capacities and delivery profiles of proteins; the specific immune response induced by PCV2 GST-ORF2-E was maintained for a relatively long period of time after immunization with the HMSN/protein complex. CONCLUSION: The findings suggest that HMSNs are good protein carriers and have high potential for use in future applications in therapeutic drug delivery.

Involvement of spinal orexin A in the electroacupuncture analgesia in a rat model of post-laparotomy pain.

BACKGROUND: Orexin A (OXA, hypocretin/hcrt 1) is a newly discovered potential analgesic substance. However, whether OXA is involved in acupuncture analgesia remains unknown. The present study was designed to investigate the involvement of spinal OXA in electroacupuncture (EA) analgesia. METHODS: A modified rat model of post-laparotomy pain was adopted and evaluated. Von Frey filaments were used to measure mechanical allodynia of the hind paw and abdomen. EA at 2/15 Hz or 2/100 Hz was performed once on the bilateral ST36 and SP6 for 30 min perioperatively. SB-334867, a selective orexin 1 receptor (OX1R) antagonist with a higher affinity for OXA than OXB, was intrathecally injected to observe its effect on EA analgesia. RESULTS: OXA at 0.3 nmol and EA at 2/15 Hz produced respective analgesic effects on the model (P<0.05). Pre-surgical intrathecal administered of SB-334867 30 nmol antagonized OXA analgesia and attenuated the analgesic effect of EA (P<0.05). However, SB-334867 did not block fentanyl-induced analgesia (P>0.05). In addition, naloxone, a selective opioid receptor antagonist, failed to antagonize OXA-induced analgesia (P>0.05). CONCLUSIONS: The results of the present study indicate the involvement of OXA in EA analgesia via OX1R in an opioid-independent way.

Numerical Analysis of Drag Reduction Characteristics of Biomimetic Puffer Skin: Effect of Spinal Height and Tilt Angle.

Based on the migratory phenomenon of the puffer and the cone-shaped structures on its skin, the effects of spinal height and tilt angle on the drag reduction characteristics is presented by numerical simulation in this paper. The results show that the trend of total drag reduction efficiency changes from slow growth to a remarkable decline, while the viscous drag reduction efficiency changes from an obvious increase to steady growth. The total and viscous drag reduction efficiencies are 19.5% and 31.8%, respectively. In addition, with the increase in tilt angle, the total drag reduction efficiency decreases gradually; the viscous drag reduction efficiency first increases and then decreases, finally tending to be stable; and the total and viscous drag reduction efficiency reaches 20.7% and 26.7%, respectively. The flow field results indicate that the pressure drag mainly originates at the front row of the spines and that the total pressure drag can be effectively controlled by reducing the former pressure drag. With the increase in low-speed fluid and the reduction in the near-wall fluid velocity gradient, the viscous drag can be weakened. Nevertheless, the drag reduction effect is achieved only when the decrement of viscous drag is greater than the increment of pressure drag. This work can serve as a theoretical basis for optimizing the structure and distribution parameters of spines on bionic non-smooth surfaces.

Regulatory T Cells in GVHD Therapy.

Graft versus host disease (GVHD) is a common complication and the leading cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Pharmacological immunosuppression used in GVHD prophylaxis and treatment lacks specificity and can increase the likelihood of infection and relapse. Regulatory T lymphocytes (Tregs) play a vital role in restraining excessive immune responses and inducing peripheral immune tolerance. In particular, clinical trials have demonstrated that Tregs can prevent and treat GVHD, without increasing the risk of relapse and infection. Hence, adoptive transfer of Tregs to control GVHD using their immunosuppressive properties represents a promising therapeutic approach. To optimally apply Tregs for control of GVHD, a thorough understanding of their biology is necessary. In this review, we describe the biological characteristics of Tregs, including how the stability of FOXP3 expression can be maintained. We will also discuss the mechanisms underlying Tregs-mediated modulation of GVHD and approaches to effectively increase Tregs' numbers. Finally, we will examine the developing trends in the use of Tregs for clinical therapy.

Shortened duration and reduced area of frozen soil in the Northern Hemisphere.

The changes in near-surface soil freeze-thaw cycles (FTCs) are crucial to understanding the related hydrological and biological processes in terrestrial ecosystems under a changing climate. However, long-term dynamics of soil FTCs at the hemisphere scale and the underlying mechanisms are not well understood. In this study, the spatiotemporal patterns and main driving factors of soil FTCs across the Northern Hemisphere (NH) during 1979-2017 were analyzed using multisource data fusion and attribution approaches. Our results showed that the duration and the annual mean area of frozen soil in the NH decreased significantly at rates of 0.13 ± 0.04 days/year and 4.9 × 104 km2/year, respectively, over the past 40 years. These were mainly because the date of frozen soil onset was significantly delayed by 0.1 ± 0.02 days/year, while the end of freezing and onset of thawing were substantially advanced by 0.21 ± 0.02 and 0.15 ± 0.03 days/year, respectively. Moreover, the interannual FTC changes were more drastic in Eurasia than in North America, especially at mid-latitudes (30°-45° N) and in Arctic regions (>75° N). More importantly, our results highlighted that near-surface air temperature (T a ) and snowpack are the main driving factors of the spatiotemporal variations in soil FTCs. Furthermore, our results suggested that the long-term dynamics of soil FTCs at the hemisphere scale should be considered in terrestrial biosphere models to reduce uncertainties in future simulations.

Exogenous Nanog alleviates but is insufficient to reverse embryonic stem cells differentiation induced by PI3K signaling inhibition.

PI3K signaling pathway plays a significant role in embryonic stem cells (ES cells) self-renewal. Overexpression of Nanog maintains mouse ES cells pluripotency independent of leukemia inhibitory factor (LIF). However, little is known about the effect of PI3K signaling pathway on ES cells with Nanog overexpression. Our experiments aimed to explore the relationship between PI3K signaling pathway and Nanog expression in ES cells. We observed the effect of LY294002, a specific inhibitor of PI3K pathway, on wild-type J1 cells and Nanog overexpressing (Ex-Nanog) J1 cells in the presence or absence of LIF. With LY294002 treatment, both of them lost their ES features even in the presence of LIF. But the differentiation induced by LY294002 on Ex-Nanog J1 cells was slighter lower than that on wild-type J1 cells. These results indicate that inhibition of PI3K pathway induces mouse ES cells differentiation. Exogenous Nanog sustains mouse ES cells pluripotency independent of LIF, and alleviates the differentiation induced by LY294002. But it is insufficient to totally reverse the differentiation.

Regulation of the class II and class I MHC pathways in human THP-1 monocytic cells by interleukin-27.

Interleukin-27 (IL-27) is an IL-12-related cytokine that can promote both anti- and pro-inflammatory immune responses. In this study, we used the promonocytic cell line THP-1, an established model for monocytes to investigate if the immunoregulatory role of IL-27 is in part due to effects on major histocompatibility complex (MHC) Ag presentation. We find that IL-27 induces mRNA and surface expression of class II MHC in THP-1 cells. IL-27 also increases class I MHC heavy chain, beta2m, and TAP-1 transcripts, leading to an increased surface expression of class I MHC. In addition, IL-27 enhances expression of costimulatory molecules CD80 and CD86 and adhesion molecule CD54. Expression of the class II transactivator (CIITA) isoforms III and IV, but not I, transcripts increases in response to IL-27. Our data suggest that the pro-inflammatory role of IL-27 is mediated in part through increased expression of key molecules involved in the class II and class I MHC pathways.

Interleukin-27 upregulates major histocompatibility complex class II expression in primary human endothelial cells through induction of major histocompatibility complex class II transactivator.

Interleukin-27 (IL-27) is a novel IL-12 family member that plays a critical role in the regulation of T-cell responses. Its immunoregulatory effects on endothelial cells (EC) remain unexplored. Here we show a role for IL-27 in the induction of major histocompatibility complex (MHC) expression in primary human EC. Stimulation of human umbilical vein ECs by IL-27 rapidly induces IFN regulatory factor-1 and dramatically increases the expression of major histocompatibility class II transactivator (CIITA) isoform IV. Expression of this transactivator correlates with increased MHC class II gene expression. IL-27 also enhances expression of MHC class I molecules. Furthermore expression of beta2-microglobulin and transporter associated with antigen processing-1 transcripts increases in response to IL-27. Additional microarray analysis demonstrates that IL-27 significantly upregulates a panel of genes that correlates with immune regulation, including the chemokines CXCL9, CXCL10, and CX3CL1 in human umbilical vein ECs. This first demonstration that both MHC II and I expression are increased in EC after IL-27 stimulation suggests that IL-27 may be important in conferring immune function on vascular endothelium.

Adipogenic potentials of mesenchymal stem cells from human bone marrow, umbilical cord and adipose tissue are different.

Mesenchymal stem cells (MSCs) could be obtained from many sources, and there are differences between them. This study was purposed to compare and analyze the basic biological characteristics of umbilical cord, adipose tissue-and bone marrow-derived MSC (UC-MSCs, AD-MSCs and BM-MSCs). The MSCs were isolated from umbilical cord, adipose tissue and bone marrow were cultured; the morphology of UC-MSCs, AD-MSCs and BM-MSCs was observed by using microscopy; the immunophenotype, differentiation potential and expression of peroxisome proliferation-activated receptor-γ (PPAR-γ) mRNA were detected by using flow cytometry, differentiation test (von kossais and 0:1 red O staining) and quantitative fluorescent PCR, respectively. The results showed that the UC-MSCs, AD-MSCs and BM-MSCs displayed similar morphology under confocal microscope after being stained with rhodamine phalloidin and DAPL. The immunophenotypes of these three originated cells conform to coincide with identification criterion for MSCs, and showed similar expression level. During adipogenic induction the adipogenic potential of these MSCs was different, AD-MSCs exhibited the highest adipogenic potential, UC-MSCs displayed the lowest, while potential of BM-MSCs get between; however, the osteogenic differentiation potential of UC-MSCs, AD-MSCs and BM-MSCs was similar. The PCR detection showed that the expression level of PPAR-γ mRNA was the highest in AD-MSCs and the lowest in UC-MSCs, while expression level in BM-MSCs get between, these results were identical with the adipogenic potential, suggest that the difference of adipogenic potential in 3 kinds of MSCs was associated with basic expression level of PPAR-γ mRNA. It is concluded that UC-MSCs, AD-MSCs and BM-MSCs exhibit similar morphology, the immunophenotypes of these MSCs coincide with identification criterion for MSCs, the osteogenic potential of these MSCs is similar, while the adipogenic potential and the expression level of PPAR-γ mRNA are different. The difference-associated mechanisms need to further study.

Stage-dependent anti-allodynic effects of intrathecal Toll-like receptor 4 antagonists in a rat model of cancer induced bone pain.

It has been reported that Toll-like receptor 4 (TLR4), which plays an important role in glial activation in neuropathic pain, is significantly increased in cancer pain. The present study was designed to assess the role of TLR4 in cancer-induced bone pain (CIBP) by intrathecal administration of TLR4 signaling pathway blocker naloxone or lipopolysaccharide Rhodobacter sphaeroides (LPS-RS). The rats developed significant mechanical allodynia from day 8 after intratibial Walker 256 inoculation. Intrathecal injection of naloxone or LPS-RS at day 8 significantly attenuated mechanical allodynia as shown by increased paw withdrawal thresholds. In contrast, the same pharmacological treatment showed no or slight pain relieving effect at day 16. Our findings demonstrate that the spinal TLR4 signaling pathway contributes to the mechanism underlying CIBP in a stage-dependent manner in rats, and it may be an efficacious target at early stage for the treatment in the future.

[Expression of human factor IX in retrovirus-transfected human umbilical cord tissue derived mesenchymal stem cells].

The purpose of this study was to investigate the expression of human Factor IX (hFIX) in retrovirus-transfected human umbilical cord tissue derived mesenchymal stem cells (hUCT-MSCs). The pLEGFP-N1-hFIX vector was generated by cloning a 3.0 kb Bgl II-BamH I fragment from the pIRES2-EGFP-hFIX plasmid containing the hFIX cDNA and part of intron 1 of hFIX in pLEGFP-N1 vector. The retroviral supernatants were produced from the Phoenix packaging cell line and then infected the hUCT-MSCs. After selection with G418 for 10 day, the expression of FIX was detected by ELISA and Western blot. The biological activity of FIX was determined by the clotting assay employing human Factor IX-deficient plasma. The results showed that compared with the activity of pooled human normal plasma (100%), transduced cells produced biologically active hFIX with 100-130% activity in two-day culture supernatant and expressed hFIX at levels of 2.68 +/- 0.36 microg/10(6) cells/24 hours after G418 selection for 10 days. The secretion of hFIX into culture supernatant was also confirmed by Western blot analysis. It is concluded that genetically modified hUCT-MSCs can express biologically active hFIX and thus serve as an efficient drug delivery vehicle carrying hFIX used as a way of somatic gene therapy for hemophilia B.

Isolation and Identification of Listeria monocytogenes from Retail Meats in Beijing.

To determine the contamination of retail meats by Listeria monocytogenes in Beijing, 70 meat samples (25 pork, 10 beef, 14 lamb, and 21 chicken) were analyzed between January and March in 1990. Eight (11%) samples were positive for L. monocytogenes , and 39 (56%) samples were found to contain other Listeria spp. Seven pork and one chicken sample contained L. monocytogenes , whereas all beef and lamb were free of L. monocytogenes . Meanwhile, 15 (60%) pork, 11 (52%) chicken, 7 (70%) beef, and 6 (43%) lamb samples were positive for other Listeria spp. The eight confirmed isolates of L. monocytogenes were serologically typed. Six (5 from pork and 1 from chicken) were serotype l/2a, and the other two isolates (2 pork) were serotype l/2c and l/2b. No isolates were of serotypes 3 and 4. All positive samples for L. monocytogenes were frozen meats, and the incidence of Listeria spp. was greater for frozen samples than for fresh. Sixty of 70 samples (22 pork, 8 beef, 11 lamb, and 19 chicken) were used to compare the effectiveness of Oxford medium and Palcam medium for the isolation of L. monocytogenes from meat samples. L. monocytogenes was isolated from seven samples by Palcam medium, while five samples were positive by Oxford medium, indicating that Palcam medium was more effective for recovering L. monocytogenes than Oxford medium.