The role and contribution of magnetic fields, characterized via their magnetic flux, to the statistical structuring of the solar atmosphere.

The anomalous heating of the solar upper atmosphere is one of the eight key problems in modern astronomy. Moreover, the stratification of the solar atmosphere is an outstanding key-problem in solar physics. In this study, a hot butterfly-like pattern is found to run through the chromosphere to the corona lying right on top of the magnetic butterfly pattern of sunspots in the photosphere. We thus propose to introduce the term butterfly body to describe the butterfly diagram in the 3-dimensional atmosphere. Besides, we discuss the so-called polar brightening in different layers. It is found to be statistically in anti-phase with the solar cycle in the photosphere and the chromosphere, while in phase with the solar cycle in the corona. Accordingly, we describe the role and relationship of solar magnetic elements of different magnetic flux strengths to explain the statistical structuring of the solar atmosphere with the butterfly body over the solar cycle.

Comparison of bacterial communities on the surface of concrete breakwater structures and ambient bacterioplankton.

Breakwater structures made of concrete are used widely around the world, and the bacteria living on these surfaces can cause the concrete to deteriorate. In this study, we collected bacterial biofilms from concrete breakwater structures located along the coast of an island, a mainland coast, and a freshwater riverbank as well as planktonic water samples from each site, and we analysed their bacterial community structures using Illumina sequencing. At the phylum level, Proteobacteria and Actinobacteria dominated planktonic samples, whereas Cyanobacteria, Proteobacteria and Bacteroidetes dominated the biofilm samples. High Cyanobacteria abundance was found in all biofilm samples. Bacterial communities significantly varied between planktonic and biofilm samples and between biofilm samples from seawater and freshwater. Only a small number of bacterial operational taxonomic units were shared by planktonic and biofilm samples from each sampling site. The permanganate index in ambient water had a more significant impact on biofilm bacterial communities than on planktonic samples. Additionally, ammonia nitrogen and total nitrogen contents were positively correlated and salinity was negatively correlated with bacterial beta diversity in biofilm samples.

Planktonic bacteria in white shrimp (Litopenaeus vannamei) and channel catfish (Letalurus punetaus) aquaculture ponds in a salt-alkaline region.

Aquaculture in salt-alkaline regions is encouraged in China, and culture of many aquatic species has been introduced into these areas. In this study, we cultured two species, white shrimp (Litopenaeus vannamei) and channel catfish (Letalurus punetaus) separately in aquaculture ponds in a salt-alkaline region in northwest China and assessed the impacts of the aquaculture operations on the planktonic bacterial community in the culture ponds. Culture of both species decreased the planktonic bacterial diversity and altered the bacterial community structure in the aquaculture ponds compared with the source water. Among the 10 dominant bacterial phyla, 8 were significantly correlated with environmental parameters; the exception was Actinobacteriota, the most dominant phylum, and Firmicutes. Proteobacteria and Bacteroidota abundances showed significant positive correlations with alkalinity, whereas Patescibacteria, Cyanobacteria, Planctomycetota, and Verrucomicrobiota abundance were positively correlated with salinity. Linear regression analysis showed that alkalinity was positively correlated with bacterial beta diversity and salinity was negatively correlated with that. In addition, white shrimp aquaculture significantly lowered the alkalinity, which suggests that culture of this species in inland salt-alkaline regions is a potential dealkalization solution.

Prednisolone dose during treatment of tuberculosis might be a risk factor for mortality in patients with systemic lupus erythematosus: a hospital-based cohort study.

Patients with systemic lupus erythematosus (SLE) are at high risk of tuberculosis (TB) because of their immunocompromised status and the use of immunosuppressive drugs. In endemic regions, TB complicates the diagnosis and treatment of SLE, but the risk factors of mortality in these patients have not been investigated. In this study, we reviewed medical records during 2006-2016. Patients who fulfilled the 1997 American College of Rheumatology SLE criteria and presented with definite TB were enrolled. The primary outcome was mortality during TB treatment. There were 5388 SLE patients screened, and 30 patients were enrolled. Seven patients died during follow-up. Compared with the survival group, patients in the mortality group had significantly more central nervous system involvement of TB, higher Systemic Lupus Erythematosus Disease Activity Index-2000 scores and more cyclophosphamide use before TB, and higher prednisolone dose before and during TB treatment. Cox regression showed that prednisolone dose during TB treatment was an independent risk factor for mortality (per 10 mg/day increase, hazard ratio (HR) 1.61, p = .019). For SLE patients, prednisolone dose during TB treatment is an independent risk factor for mortality. Keeping prednisolone dose at less than 25 mg per day during TB treatment might be a reasonable strategy in these patients.

[Analysis of risk factors for recurrent sudden sensorineural hearing loss].

Objective:To analyze the factors that may affect the recurrence of sudden sensorineural hearing loss (SSNHL).Method:We retrospectively analyzed the data of 665 SSNHL patients admitted to our hospital from January 2006 to December 2016, among which 12 patients were recurrent. The possible factors that may affect the recurrence of SSNHL were analyzed by Logistic regression analysis.Result: Hyperlipidemia might be one of the factors that caused the recurrence of SSNHL (Odds ratio 54.943,P=0.000). History of radiotherapy might be another factor that caused the recurrence of SSNHL (Odds ratio 64.100,P=0.000).Conclusion:Hyperlipidemia and the history of radiotherapy of head and neck might be risk factors for the recurrence of SSNHL.

[Research on the effect of biological glue on augmenting the viscosity of tissue engineering retinal nerve scaffolds].

Objective: To evaluate the adhesion effect of tissue engineering retinal nerve scaffolds modified with biological glue. Methods: Experimental study. To fabricate a biological glue by blending laminin, collagen Ⅳ, entactin and HSPG(2) with phosphate-buffered saline (PBS) buffer solution, scaffolds were then modified with the biological glue of various concentration levels (10 µl/cm(2), 20 µl/cm(2), 30 µl/cm(2) and 40 µl/cm(2)). The effects of various concentration levels glue on inducing scaffold adhesion were analyzed after 24 h. Cell count method and CCK-8 kit were used to assess the effects of biological glue on cell growth and toxicity. Then the scaffolds modified with or without glue were transplanted into rabbit's retina by 23 G pars plana vitrectomy. Intraocular pressure(IOP) and retina examination were assessed by ICare, fundus photograph and OCT. The adhesion effects of various concentration levels glue were analyzed by chi-square test. The cell adhesion rate of different scaffolds was analyzed by Mann-Whitney U test. One-Way ANOVA was used to compare the cell survival rate of different scaffolds. After transplantation, the IOP variance of rabbits was analyzed by repetitive measurement deviation analysis. Results: Compared with the control groups, the maximum adhesion rate of the biological glue was 91.7% (χ(2)=8.79, P<0.05) at 30µl/cm(2) concentration level. After 24h of cultivation, cell adhesion rate of glue-scaffold group (86.85%) was significantly higher than that of pure scaffold group (13.78%, U=0.01, P<0.05), while there was no significant difference between the cell survival rates of the two groups (F=7.235, P=0.11). There was no significant difference (F=79.16, P=0.07) between the IOP of viscosity modified group [(18.4+0.93) mmHg (1 mmHg=0.133 kPa)] and non- modified group [(17.1±0.04)mmHg]. The retina adhesion rate of viscosity modified scaffold was 80% at 7 days postoperatively, and the fundus examination showed no inflammatory response in retina and vitreous cavity. Conclusion: This study showed that the biological glue has favorable viscosity modifying effect on tissue engineering neural retina scaffolds, which is beneficial for the biological material transplantation. (Chin J Ophthalmol, 2018, 54: 277-282).

Decreased microRNA-146a in CD4+T cells promote ocular inflammation in thyroid-associated ophthalmopathy by targeting NUMB.

OBJECTIVE: The aim of the study was to explore the functional role of miR-146a in CD4+T cells of active thyroid-associated ophthalmopathy (TAO) patients and to identify the possible molecular mechanism for the modulation of miR-146a in TAO. PATIENTS AND METHODS: The experimental group collected six cases of peripheral venous blood of patients with active TAO. The healthy control group collected six cases of normal peripheral venous blood. All specimens excluded other eye diseases and autoimmune diseases, tumors, asthma, chronic inflammation, Human Immunodeficiency Virus (HIV), recent history of trauma, infection, and showed normal thyroid function. All patients with active TAO and age- and sex-matched healthy control subjects in this study. RESULTS: miR-146a is downregulated in active TAO CD4+T cells. NUMB was a target of miR-146a. CONCLUSIONS: We observed that miR-146a expression was downregulated in CD4+T cells during the active stage of patients with TAO. At the same time, it was found that NUMB can be targeted by miR-146a in CD4+T cells in TAO patients. Decreased microRNA-146a in CD4+T cells promotes ocular inflammation in active TAO by targeting NUMB.

[The advances in research on precisely inducing retinal ganglion cells from stem cells].

The injury and repair of retinal neurons is a common scientific problem in the occurrence, development and prognosis of neuronal visual impairment. Transplant of retinal ganglion cells (RGCs) differentiated from stem cells opens a new avenue for treatment of glaucoma and optic neuronal degenerative diseases. For the goal to explore the optimal method for RGCs replacement, this review summarizes the current information regarding the classification and application of stem cells, the growth characteristics of RGCs and the precise methods to induce RGCs, and discusses some important issues that need resolving and are related to RGCs transplantation. It is hoped that this article will provide useful theoretical basis for the research of this field. (Chin J Ophthalmol, 2017, 53: 381-385).

[Association between food intake and the serum total cholesterol level among adults in 9 regions of China].

Objective: To analyze the association between food intake and the serum total cholesterol(TC) level in adult Chinese population. Methods: This study included apparent healthy residents aged between 20 and 70 years who participated in the eighth round of China Health and Nutrition Study (CHNS) in 9 regions of China including Liaoning, Helongjiang, Shandong, Jiangsu, Henan, Hunan, Hubei, Guizhou provinces and Guangxi Zhuang Autonomous Region in 2009. The association between the serum TC level and the diet and other related factors were analyzed. Results: (1) Linear trend test showed that age, body mass index, waist circumference, systolic and diastolic blood pressure were positively related with serum TC level in 20-45 and 46-70 years old male and female (all P<0.01), while physical activity level was negatively related with serum TC level in 20-45 and 46-70 years old male and 46-70 years old female (all P<0.01). (2) Multivariate linear regression analysis showed that livestock meat intake was positively related with serum TC level in following populations: 20-45 years old male, ß=0.133, P<0.001; 46-70 years old male, ß=0.102, P=0.001; 20-45 years old male, ß=0.065, P=0.041; years old female, ß=0.059, P=0.045. Vegetable oil intake was negatively related with serum TC level in 20-45 years old male (ß=-0.071, P=0.021). Saturated fat intake was positively related with serum TC level in 46-70 years old female (ß=0.084, P=0.019). Total dietary cholesterol intake was positively related with serum TC level in male (20-45 years old, ß=0.067, P=0.021; 46-70 years old, ß=0.070, P=0.012), but not in female (20-45 years old, ß=0.007, P=0.809; 46-70 years old, ß=0.038, P=0.144). Cholesterol intake from livestock meat was positively related with serum TC level in male (20-45 years old, ß=0.156, P<0.001; 46-70 years old, ß=0.099, P=0.001), and positively related with serum TC level in 46-70 years old female (ß=0.063, P=0.028). Cholesterol intake from egg was positively related with serum TC level in 46-70 years old male (ß=0.056, P=0.032). Conclusions: Age, body mass index, waist circumference, blood pressure, blood pressure, and physical activity level are related with serum TC level. Cholesterol intake from livestock meat was the main food source that related with serum TC level, and the serum TC increases at a relatively small level after intake of total dietary cholesterol.

The influence of testicular microlithiasis on the outcomes of in vitro fertilisation in a Chinese Han population.

To investigate the influence of testicular microlithiasis (TM) on the outcomes of in vitro fertilisation (IVF), we retrospectively reviewed the ultrasonography results of the TM patients who underwent IVF treatment in our hospital. They were classified as classic-TM (CTM) or limited-TM (LTM) comparing with patients without TM (Non-TM). Semen parameters, rates of fertilisation, normal fertilisation, good embryos and clinical pregnancy were analysed. The Non-TM group showed higher percentage of morphologically normal spermatozoa than the CTM or LTM group (4.08 ± 2.07 versus 3.40 ± 2.00 versus 3.04 ± 1.66, p = .003) while the other general semen parameters were comparable. The LTM group showed higher rate of fertilisation than the CTM group (85.10% versus 81.12%, p = .044). Moreover, the rates of normal fertilisation of Non-TM (62.47%) and LTM (66.32%) group were significantly higher than the CTM (58.02%) group (p = .027 and p = .001 respectively). There were 333 embryo transfer cycles in total (including 222 fresh and 111 frozen). The overall clinical pregnancy rate was 54.95%, 58.33% and 64.12% in the group of CTM, LTM and Non-TM respectively. However, no statistically significant difference was obtained (p = .326). Our results suggest that TM may have influence on the IVF outcomes. The extent of microlithiasis correlates inversely with the rates of fertilisation and normal fertilisation.

Glucose consumption and alpha-glucosidase inhibitory activities of aqueous root extract of Helicteres angustifolia.

OBJECTIVE: The root of Helicteres angustifolia L. (Sterculiaceae) has been used as tea to treat diabetics effectively by local people in Laos. However, no scientific evidence is available for this ethnomedicinal usage. This study was undertaken to explore the hypoglycemic effect of aqueous extract from Helicteres angustifolia root. PATIENTS AND METHODS: The effect of aqueous extract from Helicteres angustifolia root on glucose consumption in C2C12 myotubes was investigated at a dose of 25, 50 and 100 µg/mL, respectively. The alpha-glucosidase inhibitory activity of the extract was evaluated using rat intestinal maltase and sucrose. Moreover, oral sucrose tolerance test (OSTT) in normal and streptozotocin induced diabetic rats was performed. Finally, their cytotoxicity in C2C12 cells and acute oral toxicity in rats was analyzed. RESULTS: Aqueous root extract of Helicteres angustifolia significantly enhanced glucose consumption in C2C12 myotubes. The extract also significantly inhibited rat intestinal maltase (IC50 = 1.44 ± 0.24 mg/mL) and sucrase activity (IC50 = 0.54 ± 0.12 mg/mL), respectively. The OSTT results showed that the extract significantly suppressed the increase of blood glucose levels in normal and diabetic rats. The extract was also proven to have low acute toxicity (LD50 > 5 g/kg) and low cytotoxicity in C2C12 cells (IC50 > 0.4 mg/mL). CONCLUSIONS: The findings from this study indicate that aqueous root extract of Helicteres angustifolia possesses significant alpha-glucosidase inhibitory activity and moderate enhanced glucose consumption activity, while with low cytotoxic and acute toxicity.

Impact of Malaria Vector Control Interventions at the Beginning of a Malaria Elimination Stage in a Dominant Area of Anopheles anthropophagus, Hubei Province, China.

Three towns with similar socio-ecological characteristics, malaria morbidities, and populations were selected for this study to explore economic and effective malaria control measures.The sources of infection were controlled in each town. Impregnated mosquito nets with 2.5% deltamethrin (15 mg/m(2)) combined with residual spraying of 5% cypermethrin (25 mg/m(2)) was implemented in cattle and pig pens, as well as in crowded sites in Chenji, whereas the mosquito nets were treated with 2.5% deltamethrin only in Guanqiao Town. All the control measures implemented in Fengling (control town) were the same as those implemented in the towns of Chenji and Guanqiao, except for mosquito elimination control. Results were evaluated and compared based on pathogens and entomology. The densities of Anopheles anthropophagus mosquitoes in houses, outside houses (man bait), as well as in cattle pens and pig pens were reduced by 100%, 71.96%, 94.01%, and 67.42%, respectively at all 4 sites in Chenji Town, whereas the density increased at 1 site (the outside house [man bait]) by 12.38%, while the densities at the other 3 sites (in houses, cattle pens and pig pens) were reduced by 99.63%, 18.71% and 69.44% respectively in Guanqiao Town. The biting rates of An. anthropophagus in the 3 towns were 0.11, 0.22, and 1.1 respectively in Chenji, Guanqiao, and Fengling. The incidence of malaria in the 3 towns decreased by 73.12%, 57.71%, and 65.71% in terms of annual average. Both impregnated mosquito nets combined with residual spraying and impregnated mosquito nets only reduced the density of An. anthropophagus in houses in the 2 towns, but reduction was more rapid in Chenji Town.

Using c-Fos/c-Jun as hetero-dimer interaction model to optimize donor to acceptor concentration ratio range for three-filter fluorescence resonance energy transfer (FRET) measurement.

Sensitized emission FRET detection method based on three-filter fluorescence microscopy is widely used and more suitable for live cell FRET imaging and dynamic protein-protein interaction analysis. But when it is applied to detect two proteins interaction in living cells, this intensity-based detection method is complicated by many experimental factors such as spectral crosstalk and spectral bleed-through and variable donor to acceptor concentration ratio. There are several FRET algorithms developed recently to correct those factors in order to quantitatively gauge and compare FRET signals between different experimental groups. But the algorithms are often difficult to choose when they are applied to certain experiments. In this research, we use c-Fos/c-Jun as a simple hetero-dimer interaction model to quantitatively detect and compare the FRET signals based on the following widely used sensitized emission FRET algorithms: N(FRET) , FRET(N) , FR, FRET(R) , E(app) and E(EFF) . We optimized the donor to acceptor concentration ratio range for the above FRET algorithms and facilitate their use in accurate FRET signal determination based on the three-filter FRET microscopy.

Increased multidrug resistance-associated protein activity in mononuclear cells of patients with systemic lupus erythematosus.

UNLABELLED: Multidrug resistance-associated proteins (MRPs, ATP binding cassette sub-family C), P-glycoprotein (P-gp) and ATP binding cassette (ABC) sub-family G member 2 (ABCG2) are important drug efflux pumps emerging after long-term medications. We intended to detect whether these molecules are expressed in immune-related cells of patients with systemic lupus erythematosus (SLE) on long-term immunosuppressants. METHODS: Mono nuclear cells (MNC) and polymorphonuclear neutrophils (PMN) were isolated from healthy volunteers and SLE patients. The MPR-mediated transport activity of these cells was measured by using carboxy-2',7'-dichlorofluorescein diacetate (CFDA) efflux assay. P-gp-mediated transport activity of cells was detected by rhodamine 123 efflux assay. ABCG2-mediated transport assay was evaluated by mitoxantrone efflux assay. The intracellular expression of MRP1, MRP2, and MRP3 molecules in MNC was detected by flow cytometry. The results were compared between MNC and PMN derived from normal and SLE groups. RESULTS: The specific dye-efflux function of MRPs in SLE-MNC is significantly higher than normal MNC. However, the expression of MRP1, MRP2, and MRP3 molecules in SLE-MNC was not different from normal MNC. We also noted that only the duration of corticosteroid treatment in different clinical/laboratory parameters was significantly correlated with the increased activity of MRPs in SLE-MNC. CONCLUSIONS: These results suggest that increased activity of MRPs in SLE-MNC is elicited by long-term corticosteroid therapy.

Abnormal in vitro CXCR2 modulation and defective cationic ion transporter expression on polymorphonuclear neutrophils responsible for hyporesponsiveness to IL-8 stimulation in patients with active systemic lupus erythematosus.

OBJECTIVE: To elucidate the molecular basis of hyporesponsiveness of polymorphonuclear neutrophils (PMN) to interleukin-8 (IL-8) stimulation in patients with active SLE. METHODS: PMN obtained from active SLE and well-matched healthy individuals were studied. The expression of two IL-8 receptors, CXCR1 and CXCR2, in PMN were detected by flow cytometry and reverse transcriptase-polymerase chain reaction. The binding affinity of PMN with IL-8 was calculated by Scatchard plotting. Soluble CXCR2 level in IL-8-stimulated PMN culture supernatant was measured by sandwich enzyme-linked immunosorbent assay. The resting and IL-8-stimulated membrane potential (MP) changes, and membrane expression of cationic ion transporters including Na+-K+-ATPase, renal epithelial Na+ channel (ENaC) and renal outer medullary epithelial K+ channel 1 (ROMK1) on PMN were detected by flow cytometry. RESULTS: Compared with normal PMN, decreased CXCR2 gene expression, but normal IL-8-binding affinity of SLE-PMN, was found. For exploring the molecular basis of the defect, the modulation of CXCR2 in SLE-PMN was intensively investigated. We found that increased cytosolic CXCR2 expression in SLE-PMN was due to defective surface translocation, increased spontaneous internalization and/or increased spontaneous synthesis. The IL-8-induced CXCR2 down-regulation in SLE-PMN was also impaired due to decreased proteolytic cleavage of IL-8-IL-8 receptor complexes from the cell surface whereas IL-8-induced internalization of the complexes was normal. In addition, we originally found that increased resting but decreased IL-8-stimulated MP in SLE-PMN was relevant to defective expression of Na+-K+-ATPase, ENaC and ROMK1 on the cell surface. CONCLUSION: The abnormal CXCR2 modulation and impaired cationic ion transporter expression cause SLE-PMN hyporesponsiveness to IL-8 stimulation in vitro.

Comparison of anti-agalactosyl IgG antibodies, rheumatoid factors, and anti-cyclic citrullinated peptide antibodies in the differential diagnosis of rheumatoid arthritis and its mimics.

OBJECTIVE: Anti-agalactosyl IgG antibodies [anti-Gal(0) IgG] have been regarded as a useful serological marker for rheumatoid arthritis (RA). Our aim was to evaluate the clinical usefulness of anti-Gal(0) IgG in the differential diagnosis of rheumatic disorders that mimic RA compared to rheumatoid factors (RF) and anti-cyclic citrullinated peptide antibodies (anti-CCP). METHODS: Sera were collected from 39 patients with RA, 49 patients with primary Sjögren's syndrome (pSjS), 47 patients with systemic lupus erythematosus (SLE), 65 patients with chronic hepatitis B viral infection (HBV), 68 patients with chronic hepatitis C viral infection (HCV) and 19 normal individuals. RF-IgM was measured by the nephelometeric method, and RF-IgA, anti-Gal(0) IgG and anti-CCP were measured by the respective ELISA assays. RESULTS: Anti-Gal(0) IgG titers were remarkably elevated in patients with RA (191.0 +/- 250.8 AU/ml) compared to pSjS (37.9 +/- 42.6 AU/ml), SLE (10.3 +/- 13.6 AU/ml), chronic HBV with (36.1 +/- 38.4 AU/ml) or without rheumatic symptoms (9.6 +/- 19.4 AU/ml), RF(+) chronic HCV without rheumatic symptoms (19.0 +/- 14.8 AU/ml), chronic HCV with rheumatic symptoms (15.2 +/- 17.4 AU/ml) and healthy individuals (2.6 +/- 0.7 AU/ml). The specificity of anti-Gal(0) IgG could be greatly enhanced by elevating the cut-off value from 12 AU/ml to 40 AU/ml (68.6% vs. 85.6%, p < 0.001) without significantly compromising its sensitivity (76.9% vs. 61.5%, p > 0.05). CONCLUSION: The serum titer of anti-Gal(0) IgG is much higher in rheumatoid arthritis than in mimicking diseases. The specificity of anti-Gal(0) IgG is enhanced when the cut-off value is raised. However, anti-CCP remains the most specific biomarker for RA.

Premature telomere shortening in polymorphonuclear neutrophils from patients with systemic lupus erythematosus is related to the lupus disease activity.

We investigated whether premature telomeric loss occurred in peripheral polymorphonuclear neutrophils (PMN) as well as mononuclear cells (MNC) from patients with systemic lupus erythematosus (SLE). We measured the telomere length of MNC and PMN in 60 SLE patients and 26 sex-, race- and age-matched healthy volunteers by Southern blotting with chemiluminescence method. The possible predisposing factors associated with telomere change were also analysed. We found the telomere length of MNC and PMN shortened with age in different degrees in both SLE and control groups. Compared to the control group, the telomere length was shortened in both SLE-MNC (6.08 kb in SLE versus 6.71 kb in control, P = 0.0008) and PMN (6.24 kb in SLE versus 6.75 kb in control, P = 0.0025). The average reduction in telomere length in SLE patients was equivalent to a premature senescence of 16.5 years in MNC and 13.4 years in PMN. In addition, the accelerated telomere shortening was more prominent in SLE patients younger than 45 years old. SLE disease activity (SLEDAI) contributed remarkably to the accelerated telomere erosion, at least in PMN. Moreover, the telomere length of MNC was significantly shorter than PMN in the same SLE patients with leukopenia and lymphopenia. These data suggested that MNC and PMN from patients with SLE displayed premature and accelerated telomere shortening that SLE is an independent factor for it.

Anti-myeloperoxidase antibodies enhance phagocytosis, IL-8 production, and glucose uptake of polymorphonuclear neutrophils rather than anti-proteinase 3 antibodies leading to activation-induced cell death of the neutrophils.

Anti-neutrophil cytoplasmic antibodies (ANCA) not only are triggered by target protein myeloperoxidase (MPO) and proteinase 3 (PR3) of polymorphonuclear neutrophil (PMN) but also react with primed PMN to exert the inflammatory process in vasculitis syndrome. To clarify the crucial role of PMN in ANCA-associated vasculitis and the related mechanism, PMN was cultured with monoclonal antibody MPO-ANCA and PR3-ANCA to determine the function of phagocytosis, Interleukin- 8 (IL-8) production, glucose uptake, and TNF-related apoptosis induced ligand (TRAIL) production. The spontaneous membrane expression of MPO and PR3 on PMN could be significantly increased by lipopolysaccharide (LPS) and TNF-alpha, but not by IL-8 or GRO-alpha. The PMN-stimulating activity of ANCA was demonstrated by enhancing phagocytosis, IL-8 production, and glucose uptake that was more prominent by MPO-ANCA. The PMN stimulation by ANCA was not through protein kinase, H2O2, or superoxide anion radicals as their inhibitors exerted no effect on ANCA-mediated activation. On the other hand, ANCA also accelerated PMN apoptosis and increased TRAIL production. These results demonstrate that activation-induced cell death (AICD) mechanism could be initiated in PMN with existence of ANCA. In conclusion, MPO-ANCA is more potent in stimulating PMN than PR3-ANCA. ANCA-activated PMN is not only responsible for the amplified inflammatory process in blood vessel but also initiates immune circuit via triggered macrophage/monocyte by apoptotic PMN through the mechanism of AICD elicited by ANCA.

Recent advances in gene expression using alphavirus vectors.

Alphavirus vectors use RNA replication in the cell cytoplasm to direct gene expression. New developments of vectors put persistency of expression and infection of specific cells in focus. Furthermore, a new application shows that the system can be used for production of retrovirus vectors carrying genes with introns and control/regulatory regions.

Packaging of intron-containing genes into retrovirus vectors by alphavirus vectors.

Efficient and controllable expression of a transgene usually requires the presence of intron sequences and much efforts have been made to produce retrovirus vectors that can transduce and integrate genes with introns. However, this has proven difficult because the viral RNA is spliced when it is synthesized in the nucleus of a producer cell. We describe a novel approach to avoid this problem. In our system the retroviral RNA is synthesized in the cytoplasm of the cell, not in the nucleus, in a reaction driven by the Semliki Forest virus (SFV) expression system. The approach was tested with a recombinant Moloney murine leukemia virus genome containing the chloramphenicol acetyltransferase (CAT) gene in association with an intron. This was inserted into a SFV transcription plasmid and the corresponding SFV vector RNA was transcribed in vitro. BHK-21 cells were then transfected with this vector RNA together with two additional SFV vectors that encode the Moloney murine leukemia virus packaging proteins. Retrovirus vectors containing intron-CAT sequences were produced at titers up to 1.3 x 10(6) infectious particles per ml during a 5-hr incubation period. The vectors faithfully transduced the intron-containing CAT gene into NIH 3T3 cells, where the intron-CAT RNA was subjected to efficient splicing and used for high level enzyme expression. Thus, the results show that intron containing genes can be efficiently packaged into retrovirus vectors by the SFV expression system.