siRNAs Derived from Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus Down-modulated the Expression Levels of Endogenous Genes in Phalaenopsis equestris.

Interplay between Cymbidium mosaic virus (CymMV)/Odontoglossum ringspot virus (ORSV) and its host plant Phalaenopsis equestris remain largely unknown, which led to deficiency of effective measures to control disease of P. equestris caused by infecting viruses. In this study, for the first time, we characterized viral small interfering RNAs (vsiRNAs) profiles in P. equestris co-infected with CymMV and ORSV through small RNA sequencing technology. CymMV and ORSV small interfering RNAs (siRNAs) demonstrated several general and specific/new characteristics. vsiRNAs, with A/U bias at the first nucleotide, were predominantly 21-nt long and they were derived predominantly (90%) from viral positive-strand RNA. 21-nt siRNA duplexes with 0-nt overhangs were the most abundant 21-nt duplexes, followed by 2-nt overhangs and then 1-nt overhangs 21-nt duplexes in infected P. equestris. Continuous but heterogeneous distribution and secondary structures prediction implied that vsiRNAs originate predominantly by direct Dicer-like enzymes cleavage of imperfect duplexes in the most folded regions of the positive strand of both viruses RNA molecular. Furthermore, we totally predicted 54 target genes by vsiRNAs with psRNATarget server, including disease/stress response-related genes, RNA interference core components, cytoskeleton-related genes, photosynthesis or energy supply related genes. Gene Ontology classification showed that a majority of the predicted targets were related to cellular components and cellular processes and performed a certain function. All target genes were down-regulated with different degree by vsiRNAs as shown by real-time reverse transcription polymerase chain reaction. Taken together, CymMV and ORSV siRNAs played important roles in interplay with P. equestris by down modulating the expression levels of endogenous genes in host plant.

Significance of M2 macrophage in tubulointerstitial disease secondary to primary Sjogren's disease.

OBJECTIVE: M2 Macrophages could improve tubulointerstitial disease in animal models. HIF-1αpromotes macrophage polarization and is involved in tubular injury. The study aims to observe the clinicopathologic significance of M2 macrophage and HIF-1α in tubulointerstitial injury secondary to primary Sjogren's disease. METHODS: Renal tissue samples from patients with tubulointerstitial disease secondary to primary Sjogren's disease (SS, n = 10), chronic tubulointerstitial nephritis secondary to drug (CIN, n = 8) were included in this study. The expression of CD163, CD68 and HIF-1α were examined by immunohistochemistry or immunofluorescence. RESULTS: (1) Renal involvement was the first manifestation in seven of ten (7/10) patients with pSS, including proteinuria, renal dysfunction, renal tubular acidosis and multiple renal stone; and two patient had intractable hypokalemia. (2) There were numerous CD163- positive cells and CD68- positive cells infiltration in tubulointerstitial injury of pSS, especially in patients with hypokalemia. CD163 positive cells and HIF-1αwere mainly expressed in acute tubulointerstitial injury of pSS, which positively correlated to N-acetyl-ß-D-glucosaminidase and ß2-microglobulin. (3) Compared with CIN, patients with pSS had higher serum globulin level, erythrocyte sedimentation rate (ESR) and lower urinary osmotic pressure. (4) During follow-up of one year, six patients with pSS and acute tubular injury acquired improved renal function on therapy of steroid and total glucosides of peony. The remaining four patients with pSS had stable renal function. CONCLUSION: M2 macrophages are involved in acute tubular injury in patients with primary Sjogren's disease. Early intervention can improve renal function of tubulointerstitial injury secondary to primary Sjogren's disease.

Recombinant adenovirus expressing a dendritic cell-targeted melanoma surface antigen for tumor-specific immunotherapy in melanoma mice model.

Viral vectors represent a potential strategy for the treatment of human malignant tumors. Currently, recombinant adenovirus vectors are commonly used as gene therapy vehicles, as it possesses a proven safety profile in normal human cells. The recombinant adenovirus system has an ability to highly express exogenous genes and increase the stability of the carrier, which is only transiently expressed in the host cell genome, without integrating. Malignant melanoma cells are produced by the skin, and melanocyte tumors that exhibit higher malignant degrees lead to earlier transfer and higher mortality. In the present study, a recombinant adenovirus (rAd) was generated to express Anti-programmed death-1 (rAd-Anti-PD-1) and used to investigate the efficacy in melanoma cells and tumors. The results demonstrated that B16-F10 cell growth was significantly inhibited and the apoptosis incidence rate was markedly promoted following rAd-PD-1 treatment. The present study demonstrated that the production of α and ß interferon was increased, which led to the induction of dendritic cell (DCs) maturation in rAd-anti-PD-1-treated mice. The present study indicated that rAd-anti-PD-1 exhibited the ability to generate more cluster of differentiation (CD)4+CD8+ T cells and induce a PD-1-specific cytotoxic T lymphocyte through DC-targeted surface antigens in mice. This resulted in a further enhanced recognition of melanoma cells due to DCs being targeted by the rAd-anti-PD-1-encoded PD-1. Notably, mice treated with the rAd-anti-PD-1-targeted PD-1 demonstrated an improved protection compared with tumor-bearing mice from the challenge group treated with a recombinant gutless adenovirus and Anti-PD-1. In conclusion, the present study demonstrated that targeting the melanoma surface antigens via the rAd-anti-PD-1-infected tumor cells enhanced the ability of recombinant adenovirus to induce a potent tumor-inhibitory effect and antigen-specific immune response.

Significance of M2 macrophages in glomerulonephritis with crescents.

OBJECTIVES: CD163 and CD206, markers of M2 macrophages, possesses anti-inflammatory properties. This study aims to investigate the clinicopathologic significance of M2 macrophages in patients of glomerulonephritis with crescents. METHODS: Renal tissue samples from patients of glomerulonephritis with more than 30% cell or cell-fibrous crescents, including lupus nephritis (LN, n=14), anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV, n=14), IgA nephropathy(IgAN) (n=11), Henoch Schonlein purpura glomerulonephritis(HSPGN)(n=8)were included in this study. The expression of CD163, CD206 and CD68 in renal tissues was detected by immunohistochemistry or immunofluorescence. RESULTS: (1) CD163 was mainly expressed in cell or cell-fibrous crescents, proliferative glomerular lesions and acute tubulointerstitial injury. There were numerous CD163-positive cells in LN and AAV in comparison with IgAN and HSPGN. (2) CD206-positive cells were mainly observed in acute tubulointerstitial injury, and proliferative glomerular lesions, especially in LN. Patients with LN had numerous CD206-positive cells in glomerular than other groups. The number of CD163-positive cells and CD206-positive cells in acute tubulointerstitial lesions of LN and AAV were more than IgAN and HSPGN. (3) Both the number of CD163-positive cells and CD206-positive cells in acute tubulointerstitial lesions negatively correlated to estimated glomerular filtration rate. (4) In LN, activity index (AI) positively correlated with the number of CD206-positive cells and CD163-positive cells. Dual staining showed that CD163-positive cells and CD206-positive cells also expressed CD68. CONCLUSIONS: CD163-positive cells and CD206-positive cells, subpopulation of macrophages, which were involved in the pathogenesis of active crescentic glomerulonephritis, especially in LN and AAV.

Age-associated potency decline in bovine oocytes is delayed by blocking extracellular Ca(2+) influx.

Oocyte aging due to delayed fertilization is associated with declining quality and developmental potential. Intracellular calcium (Ca(2+)) concentration ([Ca(2+)]i) regulates oocyte growth, maturation, and fertilization and has also been implicated in aging. Using bovine oocytes, we tested the hypothesis that oocyte aging could be delayed by reducing [Ca(2+)]ivia blocking the influx of extracellular Ca(2+) or chelating ooplasmic free Ca(2+). After IVM, cumulus-oocyte complexes or denuded oocytes were cultured in medium supplemented with 1-octanol, phorbol 12-myristate 13-acetate, or 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis-acetoxymethyl ester (BAPTA-AM) to manipulate [Ca(2+)]i. Addition of 1-mM 1-octanol increased blastocyst development rates in the cumulus-oocyte complexes aged for 6 hours by IVF and for 6, 12, and 24 hours by parthenoactivation, and this effect was independent of the presence of cumulus cells. The intracellular levels of ATP, Glutathione, and Glutathione disulfide were not affected by 1-octanol, but [Ca(2+)]i was significantly decreased. When oocytes were cultured in Ca(2+)-free medium for 12 hours, the blastocyst development rate was greater and the beneficial effects of 1-octanol on oocyte aging were abolished. However, when the medium was supplemented with phorbol 12-myristate 13-acetate, [Ca(2+)]i increased and the blastocyst development rate decreased. Moreover, BAPTA-AM reduced [Ca(2+)]i and increased blastocyst development rates after IVF or parthenoactivation. We conclude that the age-associated developmental potency decline was delayed by blocking the influx of extracellular Ca(2+) or reducing ooplasmic free Ca(2+). 1-Octanol, BAPTA-AM, or Ca(2+)-free medium could be used to lengthen the fertilization windows of aged bovine oocytes.

[Expressions of FHIT and PTEN and their significance in prostate cancer].

OBJECTIVE: To investigate the expressions of the FHIT and PTEN genes and their significance in prostate cancer. METHODS: The expressions of FHIT and PTEN were detected in 85 cases of prostate cancer and 30 cases of benign prostatic nodular hyperplasia by immunohistochemistry of PV-6000. RESULTS: The positive expression rates of FHIT and PTEN were 34.1% and 42.4% in prostate cancer, significantly lower than 96.7% and 90.0% in benign prostatic nodular hyperplasia (P <0.01). Statistically significant differences were found in the positive expression rates of FHIT and PTEN among different Gleason grades, 44.4% and 55.6% in well differentiated, 38.9% and 44.4% in moderately differentiated, and 25.0% and 37.5% in lowly differentiated prostate cancer (P <0.05). But the expression of FHIT. CONCLUSION: FHIT and PTEN may play a certain role in the was not correlated with that of PTEN in the prostate cancer tissue (P >0.05). development, progression and infiltration of prostate cancer.