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Contrasting fertilization modifies soil phosphorus (P) transformation and bioavailability, which impact crop P uptake and P migration in the soil profile. A long-term (25-year) fertilizer experiment was employed to investigate crop yield, P uptake and changes in sequentially extracted P fractions in the soil profile, and their relationships on a calcareous soil derived from loess material under a winter wheat and summer maize double-cropping system. The experiment involved seven nutrient management treatments: control (CK, no nutrient input), N, NK, NP, and NPK, representing various combinations of synthetic nitrogen (N), phosphate (P), and potassium (K) applications, as well as combinations of NPK fertilizers with either crop residues (SNPK, where S refers to maize stalk or wheat straw) or manure (MNPK, where M refers to dairy manure). Wheat and maize yields were significantly higher with P input fertilizer relative to the P-omitted treatments. Long-term application of P-containing fertilizers markedly raised the contents of inorganic (Pi) and organic (Po) P fractions at 0–20 cm depth compared with the P-omitted treatments. Moreover, both Pi and Po fractions were markedly higher under MNPK than under NPK and SNPK treatments. For achieving high yield for wheat and maize, the critical contents of labile P were 54 and 63 mg kg−1, and those of moderately labile P were 48 and 49 mg kg−1, respectively, defined by the linear plateau model. In addition, the change points of labile P and moderately labile P were 99 and 70 mg kg−1, above which CaCl2-P content significantly increased. Moreover, long-term P input significantly accumulated different P fractions in the deeper soil layers up to 100 cm, with large portions of organic P being a composite of labile and moderately labile P, especially in MNPK treatment. Our results suggest that excessive P supply with organic manure resulted in massive P accumulation in the topsoil and promoted soil P fraction transformation and availability in the deep soil layers, especially in an organic P form that has often been neglected.

Abstract Objective Neutrophils and aberrant NETosis have been implicated in the pathogenesis of diverse autoimmune diseases; however, their roles in primary Sjögren’s syndrome (pSS) remain unclear. We aimed to reveal the potential roles of neutrophils and neutrophil extracellular traps (NETs) in pSS. Methods pSS patients were enrolled and NETosis markers were measured in plasma and labial glands using ELISA and immunofluorescence. The gene signatures of neutrophils were assessed by RNA-Seq and RT-PCR. Reactive oxygen species (ROS), mitochondrial ROS (MitoSOX) production, and JC-1 were measured by flow cytometry. Results NETosis markers including cell-free DNA (cf-DNA) and myeloperoxidase (MPO) in plasma and labial glands from pSS patients were significantly higher than healthy controls (HCs) and were associated with disease activity. RNA sequencing and RT-qPCR revealed activated type I IFN signaling pathway and higher expression of genes related to type I interferon in pSS neutrophils. Further stimulating with IFN-α 2a in vitro significantly induced ROS production and JC-1 monomer percentage in pSS neutrophils. Conclusions Our data suggest the involvement of neutrophils and enhanced NETosis in pSS patients. Further mechanism study in vitro revealed that type I IFN activation in pSS neutrophils led to mitochondrial damage and related ROS production which finally result in the generation of NETs.

Partial substitution of synthetic nitrogen fertilizer with manure (organic substitution) is highly recommended to minimize environmental risks without compromising crop productivity in intensive agricultural systems. However, our understanding of the effect of organic substitution on soil organic carbon (OC) and total nitrogen (TN) in deep soil and its impact on crop productivity remains limited. Here we investigated OC and TN changes in soil profile down to 100 cm, crop yield, and sustainable yield index under synthetic nitrogen, phosphate, and potassium fertilizers (NPK), NPK plus straw (NPKS), and organic substitution (NPKM) treatments over two decades in four upland fields across different climate zones. Compared with the initial values, two decades of NPKM treatment significantly (p < 0.05) increased OC and TN stocks in either topsoil (by 25.6–103.8 and 15.8–89.8%) or deep soil (by 2.9–71.3 and 5.7–36.9%), respectively, across all sites. The increases in OC and TN stocks in 0–100 cm soil receiving NPKM were significantly higher than those receiving NPK at all sites and NPKS at three high-evaporation sites. Compared with NPKS and NPK treatments, crop yield and N uptake were significantly increased under NPKM treatment only at the Qiyang site. Furthermore, OC sequestration in the entire soil profile down to 100 cm and TN accumulation in topsoil exhibited significant positive correlations with crop N uptake, relative crop yield, and sustainable yield index. In conclusion, long-term partial substitution of synthetic N fertilizer with manure facilitates soil OC and TN sequestration in the entire 100 cm profile and thus maintains high crop productivity in upland areas.

BackgroundsObservational studies have identified associations between smoking, alcohol use, body mass index (BMI), and the levels of vitamin D with primary biliary cholangitis (PBC). However, there was a lack of randomization control studies to estimate the causal relationship. This study was to investigate the causal estimates for the effects of those risk factors on PBC.MethodsThe genetic instrument variants were extracted from genome-wide association studies in European ancestry. Two-sample mendelian randomization (MR) and multivariable mendelian randomization were used to determine genetically causal estimates. Primary analyses consisted of random-effects and fix-mode inverse-variance-weighted methods, followed by secondary sensitivity analyses to verify the results.ResultsOur study showed that BMI was a causal factor for PBC (OR 1.35; 95% CI=1.03-1.77; p=0.029). In addition, we found that serum vitamin D levels had a protective effect on PBC after adjusting for BMI (OR 0.51; 95% CI=0.32-0.84; p=0.007). However, we failed to identify evidence supporting that genetic causal effect of smoking and alcohol intake were associated with PBC in European countries.ConclusionOur results enriched findings from previous epidemiology studies and provided evidence from MR that serum vitamin D concentrations and BMI were independent causal factors for PBC, suggesting that ensuing vitamin D sufficiency and healthy lifestyles might be a cost-effective measure for early intervention for PBC.

BackgroundProgrammed cell death protein-1/programmed cell death ligand-1 (PD-1/PD-L1) inhibitors therapy is now a routine scheme in cancers. However, the effect of preexisting autoantibodies on the safety and efficacy of PD-1/PD-L1 inhibitors in cancer patients is not well understood.MethodsThe present retrospective cohort study evaluated the safety and efficacy of PD-1/PD-L1 inhibitors in patients with preexisting autoantibodies. Patients who received PD-1/PD-L1 inhibitors in the Department of Medical Oncology, Peking Union Medical College Hospital between November 2017 and August 2021 were reviewed.Results67 (37.9%) of the 177 patients, 27 (20.3%) of the 133 patients, and 16 (11.0%) of 146 patients who received PD-1/PD-L1 inhibitors were positive for ANA, anti-Ro52, and antithyroid antibodies, respectively. Preexisting ANA and anti-Ro52 antibody were not associated with the increased risk of immune-related adverse events (irAEs), while thyroid dysfunction was more frequent in patients with positive antithyroid antibody (75.0% versus 13.8%, p < 0.001). The median progression-free survival (PFS, 13.1 versus 7.0 months, p = 0.015) was significantly longer in the ANA-positive patients, while the median overall survival (OS, 14.5 versus 21.8 months, p = 0.67) did not differ significantly between the ANA-positive and ANA-negative groups. Moreover, the preexisting anti-Ro52 and antithyroid antibodies were not significantly associated with PFS and OS.ConclusionsThe presence of ANA and anti-Ro52 antibody were not associated with a higher risk of irAEs, whereas patients positive for antithyroid antibody should monitor closely immune-related thyroid dysfunction. Preexisting ANA might be a predictor of longer PFS, while anti-Ro52 and antithyroid antibodies had no significant effect on survival outcomes in patients receiving PD-1/PD-L1 inhibitors therapy.

Here we reported two anti-Mi-2 autoantibody-positive dermatomyositis (DM) patients with a characteristic antinuclear antibody (ANA) immunofluorescence pattern. Autoantibodes were screened by indirect immunofluorescence (IIF) on HEp-2 cells (Euroimmun, Lübeck, Germany) and confirmed by line immunoblot (ANA Profile 3—Euroimmun, Germany). These two patients were positive for ANA (speckled, titer 1:320), followed by confirmation of positive anti-Mi-2α and anti-Mi-2β positive and negative for all other antibodies. We found a characteristic ANA pattern of the anti-Mi-2 antibody that differed from the AC-4 pattern, especially in the morphology of mitotic cells (metaphase, anaphase, and telophase). Thus, we would like to suggest reporting this characteristic antinuclear antibody pattern as a new AC type, as AC-X.

Seeking food security, contemporary Chinese agriculture has followed a trajectory of overfertilization and associated environmental problems, hence the need for nitrogen-balancing practices that do not compromise yield and quality. Here we present a national meta-analysis using 224 studies with 1972 comparisons to quantify the potential to reduce nitrogen (N) fertilization to improve environmental outcomes while maintaining yield and grain protein. We calculated a nitrogen reduction ratio (NRR), as 100 × ( N _C − N _T )/ N _C ; where N is N fertilizer rate and subscripts indicate farmer practice (C) and reduced N rate treatment (T). Our meta-analysis showed that the NRR that maintained yield and grain protein content at the level of current practice was up to 10% in wheat and up to 30% in maize and rice. Larger yield-neutral NRR could be achieved in more fertile, heavier-textured soils, and with practices including enhanced-efficiency N fertilizer, combined application of organic and inorganic N fertilizer, and incorporated straw. Assuming a reduction in N fertilizer usage by 10% for wheat and by 30% for maize and rice in the current cropping area, there is a potential to save 5.7 Mt N yr ^−1 ; reduce loss of reactive nitrogen by 1.26 Mt N yr ^−1 , equivalent to 63% of annual total Nr losses for rice in China, reduce N-related greenhouse emissions by 75.2 Mt CO _2 -eq yr ^−1 , equivalent to 14.5%–25% of the emissions associated with the N fertilizer chain in China; and improve N use efficiency by 23%. Our results highlight the feasibility of maintaining yield and grain protein, and achieving substantial environmental benefits with reduced fertilization rate, and the environmental and agronomic scenarios where these outcomes are more likely.

Plastic film mulching often increases the yield of winter wheat in the Loess Plateau of China, but the physiological mechanisms are unclear, especially in response to the interaction between mulch and water supply. In this study, we investigated the interactive effects of initial soil water (dry, moderate, and wet), soil cover (plastic mulch, bare soil), and seasonal conditions on the dynamics of dry matter partitioning, water-soluble carbohydrates (WSC), and flag leaf senescence, and their relations with yield and its components. Plastic mulch increased dry matter accumulation at anthesis and maturity relative to bare soil, with no interaction with season or initial soil water. Allocation of dry matter to leaf, stem, and spike did not change with soil cover. Compared with bare soil, mulch increased WSC accumulation by 14% at anthesis and its translocation by 16%. Soil cover did not influence the senescence of flag leaf after anthesis as indicated by similar dynamics of the C:N ratio. Grain yield was higher under plastic mulch than bare soil in two out of three seasons, and was associated with a higher translocation amount of WSC and post-anthesis dry matter that linked grain weight, grain number, and harvest index.

Organic phosphorus (Po) may play a vital role in phosphorus availability via its mineralization by alkaline phosphatase (ALP), being encoded by phoD gene, in calcareous soil. Understanding the effects of long-term fertilization on the community of phoD harboring bacteria and the related alteration of the P availability owing to the changes in ALP secretion may offer a chance to elucidate the Po contribution to soil available P. Based on a long-term experiment, we analyzed the phoD gene harboring microbial diversity, abundance and composition, ALP and Po forms, and their relationship. The treatments involved were control without any fertilizers (CK), synthetic nitrogen and potassium (NK), synthetic nitrogen, phosphorus and potassium (NPK), NPK and crop stalk return (SNPK), and NPK plus organic manure (MNPK). Fertilization increased the abundance and diversity of phoD gene harboring microbial over CK. Those receiving NPK and NPK treatments integrated with organic supplements significantly improved the relative abundance of Proteobacteria but decreased Gemmatimonadetes at the phylum level, while all fertilized treatments appreciably increased the relative abundance of Lysobacter but decreased that of Gemmatirosa and Afipia, at the genus level. SNPK and MNPK treatments noticeably increased the relative abundance of Methylobacter but reduced Pseudomonas and Streptomyces relative to those receiving synthetic fertilizer treatments. Long-term fertilization markedly raised ALP activity, which was significantly and positively correlated with the relative abundance of the phylum Proteobacteria as represented by the genera Methylobacterium and Lysobacter. ALP was closely associated with moderately labile Po, followed by enzyme P, recalcitrant Po, and labile Po. The changes in phoD bacteria and ALP were mainly driven by soil organic carbon, Olsen P and pH. We concluded that the long-term fertilization, especially the addition of organic supplements, profoundly modified the soil properties and subsequently changed the diversity and relative abundance of phoD gene harboring bacteria, which promoted the activity of ALP, and thus the mineralization of various forms of Po (mainly moderately labile Po) to enhance the P availability.

Abstract Background Ovarian cancer (OC) is a common gynecological cancer and characterized by high metastatic potential. MicroRNAs (miRNAs, miRs) have the promise to be harnessed as prognostic and therapeutic biomarkers for OC. Herein, we sought to identify differentially expressed miRNAs and mRNAs in metastatic OC, and to validate them with functional experiments. Methods Differentially expressed miRNAs and mRNAs were screened from six pairs of primary OC tissues and metastatic tissues using a miRStar™ Human Cancer Focus miRNA and Target mRNA PCR Array. Then, gene expression profiling results were verified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot assays. The binding affinity between miR-7-5p and TGFβ2 was validated by dual-luciferase reporter assay. Expression of miR-7-5p and TGFβ2 was manipulated to assess their roles in malignant phenotypes of highly metastatic HO-8910PM cells. Results MiRNA profiling and sequencing identified 12 miRNAs and 10 mRNAs that were differentially expressed in metastatic tissues. Gene ontology and Pathway analyses determined that 3 differentially expressed mRNAs (ITGB3, TGFβ2 and TNC) were related to OC metastasis. The results of RT-qPCR confirmed that the decrease of miR-7-5p was most significant in OC metastasis, while TGFβ2 was up-regulated in OC metastasis. Moreover, miR-7-5p targeted and negatively regulated TGFβ2. MiR-7-5p overexpression accelerated HO-8910PM cell viability and invasion, and TGFβ2 overexpression reversed the results. Meanwhile, simultaneous miR-7-5p and TGFβ2 overexpression rescued the cell activities. Conclusions This study characterizes differentially expressed miRNAs and mRNAs in metastatic OC, where miR-7-5p and its downstream target were most closely associated with metastatic OC. Overexpression of miR-7-5p targets and inhibits TGFβ2 expression, thereby inhibiting the growth and metastasis of OC.

The root system of a plant provides vital functions including resource uptake, storage, and anchorage in soil. The uptake of macro-nutrients like nitrogen (N), phosphorus (P), potassium (K), and sulphur (S) from the soil is critical for plant growth and development. Small signaling peptide (SSP) hormones are best known as potent regulators of plant growth and development with a few also known to have specialized roles in macronutrient utilization. Here we describe a high throughput phenotyping platform for testing SSP effects on root uptake of multiple nutrients. The SSP, CEP1 (C-TERMINALLY ENCODED PEPTIDE) enhanced nitrate uptake rate per unit root length in Medicago truncatula plants deprived of N in the high-affinity transport range. Single structural variants of M. truncatula and Arabidopsis thaliana specific CEP1 peptides, MtCEP1D1:hyp4,11 and AtCEP1:hyp4,11, enhanced uptake not only of nitrate, but also phosphate and sulfate in both model plant species. Transcriptome analysis of Medicago roots treated with different MtCEP1 encoded peptide domains revealed that hundreds of genes respond to these peptides, including several nitrate transporters and a sulfate transporter that may mediate the uptake of these macronutrients downstream of CEP1 signaling. Likewise, several putative signaling pathway genes including LEUCINE-RICH REPEAT RECPTOR-LIKE KINASES and Myb domain containing transcription factors, were induced in roots by CEP1 treatment. Thus, a scalable method has been developed for screening synthetic peptides of potential use in agriculture, with CEP1 shown to be one such peptide.

Background: The liver and skin are the most common organs involved in Stevens–Johnson syndrome/toxic epidermal necrolysis (SJS/TEN). Drug reactions rarely affect both organs concurrently. The clinical features, risk factors, and prognostic markers of drug-induced liver injury (DILI) in patients with SJS/TEN are not well studied. Materials and Methods: The clinical features, risk factors, and prognostic markers of DILI in patients with SJS/TEN hospitalized at the dermatology department of our hospital from January 2009 to December 2018 were retrospectively analyzed. Results: A total of 298 patients with SJS/TEN were enrolled in this study. Of them, 40 had liver injury and the rest served as control. Causative drugs mainly included antipodagrics (xanthine oxidase inhibitors occupying 100% among antipodagrics), anticonvulsants (dibenzazepine occupying 76.92% among anticonvulsants), and traditional Chinese medicines. There was a statistically significant difference between the patients with liver injury and the control group in the history of liver disease, diabetes, and hyperlipidemia (P < 0.05). Nine of the 40 patients with liver injury died. High serum total bilirubin and creatinine levels were significantly associated with poor prognosis of DILI in patients with SJS/TEN (P < 0.05). Conclusion: DILI usually occurs in patients with SJS/TEN. Pre-existing liver disease, diabetes, and hyperlipidemia are independent risk factors for DILI in patients with SJS/TEN. High serum total bilirubin and creatinine levels may be useful prognostic markers for DILI in patients with SJS/TEN.

The combination of organic manure and inorganic fertilizer plays a role in increasing crop yield and nutrient efficiency, but such effectiveness varies with crop, soil, management, and climate. Here, a 6-year field experiment was conducted to evaluate the effects of substituting organic manure with inorganic fertilizer on crop yield, grain protein content, and nitrogen and phosphorus efficiency under a winter wheat-summer maize cropping system on Anthrosol. Five treatments were included: recommended nitrogen (N), phosphorus (P) and potassium (K) fertilizers (NPK), 75% NPK + 25% organic manure (M), 50% NPK + 50% M, 25% NPK + 75% M, and 100% M, respectively. Wheat, maize, and annual yield were 1643–8438 kg ha−1, 4847–11,104 kg ha−1, and 10,007–17,496 kg ha−1. Organic manure treatments produced the same crop yield as NPK treatment except for 100% M. Grain protein content of wheat and maize was 7.9–15.1% and 5.6–12.6%. Organic manure treatments yielded significantly lower wheat grain protein content but had no significant effect on maize grain protein content relative to NPK treatment. Nitrogen uptake efficiency and nitrogen use efficiency at the cropping system level were 0.67–1.16 and 35.7–60.5 kg kg−1. Phosphorus uptake efficiency and phosphorus use efficiency were 0.28–0.75 and 167–531 kg kg−1. Compared with NPK treatment, 50% M, 75% M, and 100% M improved nitrogen use efficiency but decreased nitrogen uptake efficiency and phosphorus efficiencies. Annual N and P budgets were −1.3–79.1 kg ha−1 a−1 and 25.6–100.1 kg ha−1a−1, and both increased with the increase in organic manure input. Based on crop yield, grain protein content, nitrogen, and phosphorus efficiency and their budget, substitution of 25% inorganic fertilizer with organic manure is the rational combination under the winter wheat–summer maize system on an Anthrosol.

Biostimulants can enhance horticultural crop production. However, their application in hydroponically grown lettuce is still limited, and information regarding their relative efficacy is lacking. A greenhouse trial was conducted to address this issue. Five nutrient solution treatments were evaluated on two lettuce cultivars: butterhead and red oak-leaf. The treatments included a half-strength modified Hoagland solution (Hs-H); a full-strength modified Hoagland solution (Fs-H); and Hs-H supplemented with 50 mg L−1 fulvic acid (FA), 334 mg L−1 seaweed extract (SE), or 5 mL L−1 gamma polyglutamic acid (PGA). The results indicated that the shoot biomass observed after biostimulant supplementation was significantly greater than or comparable to that observed with Fs-H. Nutrient solutions supplemented with SE and PGA led to a greater increase in the root biomass than that realized with Hs-H and Fs-H treatments. The Hs-H + FA treatment resulted in the lowest root-to-shoot ratio on a fresh weight basis among all treatments. The nitrate concentration in the shoot was significantly reduced following biostimulant supplementation compared to that realized with Fs-H and Hs-H treatments. Nutrient solutions supplemented with SE and PGA also decreased soluble sugar concentrations compared to that achieved using Hs-H and Fs-H treatments. FA and SE improved nutrient uptake for both cultivars, but PGA had a minimal effect on nutrient uptake. The two cultivars varied in their responses to biostimulant supplementation with regard to biomass, quality traits, and nutrient uptake. This study supports using fulvic acid and seaweed extract, rather than γ-PGA, in hydroponic lettuce production systems.

Temporal changes in the concentrations of plant-available phosphorus (P) in soil (Olsen-P), total soil-P and P activation coefficient (the ratio of Olsen-P to residual-P (i.e. an approximation to total-P)) were measured in plots that received consistent inorganic nitrogen, phosphorus and potassium plus organic fertilizers annually. Maize and winter wheat crops were grown in rotation for 24 years. Olsen-P and P activation coefficient declined significantly in the earlier years (< 12 years) for treatments that did not include any P fertilizer, and increased over the same period for the P-fertilized treatments. The rates of change in the Olsen-P and P activation coefficient values were positively related to P balance. In the later years, the Olsen-P and P activation coefficient plateau values were positively related to the P balance.

Abstract Objective To determine the adult reference intervals for the Siemens IgG subclass reagents. Methods 636 blood samples of healthy adults were analyzed to determine the level of IgG subclass using the reagents of Siemens immunonephelometric assay with molecular biology kits. Results IgGSc reference intervals were as follows: IgG1 4.45–9.76 g/L, IgG2 2.07–8.57 g/L, IgG3 0.08–0.80 g/L and IgG4 0.05–1.54 g/L. There was an excellent correlation between the total IgG and the sum of the IgG subclasses. No significant gender and age differences were observed. Conclusions Our data provide the missing reference intervals and enable the use of the nephelometric IgG subclass reagents in Chinese. The study can offer reference on clinic diagnose.

Abstract Detection of autoantibodies in systemic lupus erythematosus (SLE) plays an important role in timely diagnosis and earlier treatment of SLE. In this study, we used a SmD1 polypeptide-based ELISA to determine anti-SmD1 antibody in 269 SLE, including100 naïve (had not been treated with steroids or immunosuppressants at study inception) SLE patients and 169 non-naive SLE patients; 233 controls with other rheumatic diseases (RDC) (70 RA, 40 AS, 73SSc, and 50 SS), and 110 healthy controls (HC) group. The positive rate of anti-SmD1 among all SLE patients was 60.97%, higher than that in the RDC group (13.30%, P = 0.000) or the HC group (9.09%, P = 0.000). The positive rate of anti-SmD1 in non-naive SLE patients was higher than that for anti-dsDNA antibodies (44.97%, P = 0.03). Positivity for anti-SmD1 only was found in 14.00% of naive SLE patients and 16.00% of non-naive SLE patients. In naive SLE patients, the serum concentration of anti-SmD1 was lower after treatment than before treatment (P = 0.039). Active SLE patients positive for anti-SmD1 were more likely to have malar rash, rash, nonscarring alopecia, PAH and hypocomplementemia. High positivity for anti-SmD1 only in patients with SLE indicated the importance and necessity of detection of anti-SmD1 in patients with SLE.

Abstract Background Dickkopf-4 (DKK4), a member of DKK family, appears to be a divergent protein. It remained multi-biological functions in carcinogenesis. The effect of DKK4 on the ovarian cancer cells remains unclear. This study detected the clinical significance of DKK4 in epithelial ovarian cancer (EOC) patients and its role in invasion. Methods QRT-PCR and western blot analysis were used to examine the levels of DKK4 mRNA and protein in 33 EOC tissues and 33 benign ovarian tumors. Immunohistochemical analysis was performed to assess DKK4 expression in 239 EOC samples. siRNA-mediated DKK4 silence was conducted. Transwell assay was used to detect the invasive ability. Phalloidin was used to stain the formations of actin filaments. Results The expressions of DKK4 mRNA and protein were elevated in EOC tissues as compared with those in benign ovarian tumors (p = 0.001 and

To understand the mechanisms affecting water balance partitioning during fallow on drylands could improve the fallow management practices in arable land ecosystems. A three-year field experiment was conducted to evaluate the effects of field management regimes on water balance partitioning and fallow efficiency during the fallow periods under a winter wheat (Triticum aestivum L.) fallow system on the Loess Plateau, China. The fallow management regimes tested were: (i) conventional practice, (ii) catch cropping, and (iii) no tillage with wheat straw mulching. A process-oriented ecosystem model (CoupModel) was calibrated with field measurements and then used to generate comparative simulations of the water balance partitioning. The simulations indicated that mulching increased the soil water storage change by 38-71 mm during the three fallow periods, thus resulting in higher fallow efficiency by 9-12%, and decreased soil evaporation by 22-72 mm, compared with the conventional practice. Furthermore, water reached deeper horizons, resulting in 7 mm deep percolation in a wet year under mulching but not under conventional practice or catch cropping. The simulation results also showed that the catch cropping decreased the soil water storage change by 13-21 mm, although it lowered soil evaporation by 11-51 mm, and altogether reduced the fallow efficiency by 3-9%, compared to conventional practice. On the Loess Plateau of China mulching proved to be a sound measure for ensuring certain fallow efficiency and possibly benefit to the water cycle, while catch cropping negatively partitioned the water balance. The catch cropping under mulching might be another management regime to be considered.

Background/Aims: The aggressive manner of ovarian cancer (OVC) cells accounts for the majority of its lethality. Recently, we have shown that placental growth factor (PLGF) promotes metastases of OVC cells through miR-543-regulated MMP7. In the current study, we analyzed the effects of PLGF on another cell invasion associated protein, ZEB2, in OVC cells. Methods: The PLGF and ZEB2 levels in OVC tissues were compared to the paired adjacent non-tumor ovary tissue. We modified ZEB2 levels in OVC cells, and examined its effects on PLGF mRNA and protein levels by RT-qPCR and by Western blot, respectively. We also modified PLGF levels in OVC cells, and examined its effects on ZEB2 mRNA and protein levels by RT-qPCR and by Western blot, respectively. Then, we examined the cell invasiveness in PLGF-modified OVC cells in a transwell cell invasion assay. Finally, we used specific signal pathway inhibitors to treat PLGF-modified OVC cells and examined the effects on ZEB2 activation. Results: PLGF and ZEB2 levels were both significantly increased in OVC tissues, compared to the paired adjacent non-tumor ovary tissue. The PLGF and ZEB2 levels were strongly correlated. ZEB2 modification did not alter PLGF levels. Overexpression of PLGF in OVC cells significantly increased ZEB2 levels and cell invasiveness, while PLGF depletion in OVC cells significantly decreased ZEB2 levels and cell invasiveness. Application of a specific MAPK-p38 inhibitor, but not application of specific inhibitors for MAPK-p42/p44, PI3k/Akt, or JNK signaling pathways, to PLGF-overexpressing OVC cells substantially abolished the PLGF-induced ZEB2 activation. Conclusion: PLGF enhances OVC cell invasion through MAPK-p38-dependent activation of ZEB2.

Background/Aims: Elucidation of the molecular mechanisms underlying ovarian cancer (OC) cell invasion and migration may provide important evidence for developing efficient therapy. Recently, Matrix metalloproteinase (MMP) has been shown to be regulated by vascular endothelial growth factor family members, especially placental growth factor (PLGF), as a potential mechanism underlying cancer invasion. Here, we studied the molecular relationship between PLGF and MMP7 in the OC. Methods: We examined the levels of PLGF and MMP7 in the resected OC specimens and compared to paired adjacent non-tumor ovarian tissue. We also examined the correlation between PLGF and MMP7. We modified PLGF levels in a human OC cell line, OVCAR3, and analyzed the effects on MMP7. Prediction of microRNA (miRNA) binding to 3'-UTR of MMP7 mRNA was performed by bioinformatics analyses and confirmed by a dual luciferase reporter assay using miR-543-modifed OC cells. The levels of miR-543 were examined in the OC specimens, and the correlation between miR-543 and PLGF or MMP7 was performed. Results: PLGF and MMP7 both significantly increased in the OC specimens, compared to paired adjacent non-tumor ovarian tissue. PLGF significantly increased MMP7 in the OC cells at protein level, but not at mRNA level. In OC cells, PLGF significantly decreased the levels of miR-543, which suppressed the translation of MMP7 mRNA via 3'-UTR binding. In OC specimen, miR-543 significantly decreased, compared to paired adjacent non-tumor ovarian tissue. An inverse correlation was detected between the levels of miR-543 and PLGF or MMP7 in the OC specimens. Conclusion: MiR-543 inhibits translation of MMP7 through binding to the 3'-UTR of MMP7 mRNA in OC. PLGF suppresses miR-543, which activates MMP7-mediated cancer invasion.

[This corrects the article DOI: 10.1371/journal.pone.0089350.].

A reference interval (RI) for the circulating concentration of anti-dsDNA antibody is essential for clinicians to interpret laboratory results and make clinical decisions. Therefore, we aimed to establish the RI for anti-dsDNA antibody in the Chinese Han population.This study was designed and carried out in accordance with guideline C28-A3, which is proposed by the International Federation of Clinical Chemistry and the Clinical and Laboratory Standards Institute. A total of 2,880 apparently healthy individuals were enrolled using a posteriori sampling. These individuals were recruited from four hospitals, representing the Han populations of north, south, east, and west China. Serum anti-dsDNA antibody levels were measured using the three analytical systems AESKU, EUROIMMUNE, and INOVA, which are the most commonly used systems in China. Individuals were stratified by gender, age, and region, and the RIs were obtained by nonparametric methods.Gender-specific RIs for serum anti-dsDNA antibody in the Chinese Han population were established.This is the first exploration of the RI for anti-dsDNA antibody in the Chinese Han population. We have established gender-specific RIs for each assay method commonly used in China.

We investigated the significance of high- mobility group box1 (HMGB1) and T-cell-mediated immunity and prognostic value in cervical cancer. HMGB1, forkhead/winged helix transcription factor p3 (Foxp3), IL-2, and IL-10 protein expression was analyzed in 100 cervical tissue samples including cervical cancer, cervical intraepithelial neoplasia (CIN), and healthy control samples using immunohistochemistry. Serum squamous cell carcinoma antigen (SCC-Ag) was immunoradiometrically measured in 32 serum samples from 37 cases of squamous cervical cancer. HMGB1 and SCC-Ag were then correlated to clinicopathological characteristics. HMGB1 expression tends to increase as cervical cancer progresses and it was found to be significantly correlated to FIGO stage and lymph node metastasis. These findings suggest that HMGB1 may be a useful prognostic indicator of cervical carcinoma. In addition, there were significant positive relationships between HMGB1 and FOXP3 or IL-10 expression (both p < 0.05). In contrast, HMGB1 and IL-2 expression was negatively correlated (p < 0.05). HMGB1 expression may activate Tregs or facilitate Th2 polarization to promote immune evasion of cervical cancer. Elevated HMGB1 protein in cervical carcinoma samples was associated with a high recurrence of HPV infection in univariate analysis (p < 0.05). HMGB1 expression and levels of SCC-Ag were directly correlated in SCC (p < 0.05). Thus, HMGB1 may be a useful biomarker for patient prognosis and cervical cancer prediction and treatment.

Resistin is a novel hormone that is secreted by human adipocytes and mononuclear cells and is associated with obesity, insulin resistance and inflammation. Recently, resistin has been postulated to play a role in angiogenesis. Here, we investigated the hypothesis that resistin regulates ovary carcinoma production of vascular endothelial growth factor (VEGF) and the angiogenic processes. We found that in human ovarian epithelial carcinoma cells (HO-8910), resistin (10–150 ng/mL) enhanced both VEGF protein and mRNA expression in a time- and concentration-dependent manner, as well as promoter activity. Furthermore, resistin enhanced DNA-binding activity of Sp1 with VEGF promoter in a PI3K/Akt-dependent manner. PI3K/Akt activated by resistin led to increasing interaction with Sp1, triggering a progressive phosphorylation of Sp1 on Thr453 and Thr739, resulting in the upregulation of VEGF expression. In an in vitro angiogenesis system for endothelial cells (EA.hy926) co-cultured with HO-8910 cells, we observed that the addition of resistin stimulated endothelial cell tube formation, which could be abolished by VEGF neutralizing antibody. Our findings suggest that the PI3K/Akt-Sp1 pathway is involved in resistin-induced VEGF expression in HO-8910 cells and indicates that antiangiogenesis therapy may be beneficial treatment against ovarian epithelial carcinoma, especially in obese patients.

Objective: To measure Lewis y and integrin α5β1 expression in epithelial ovarian carcinoma and to correlate the levels of these molecules with ovarian carcinoma chemotherapy and prognosis. Methods: The study population included 34 ovarian carcinoma patients with chemotherapeutic drug-resistance, six partially drug-sensitive cases, and 52 drug-sensitive cases (92 total). Immunochemistry was used to determine expression of Lewis y antigen and integrin α5β1 in ovarian carcinoma tissues, and correlation of these molecules with chemotherapy resistance was further investigated, Multi-factor logistic regression analysis was applied to investigate: age, surgical stage, grade, subtype of patient cases, metastasis of lymph nodes, residual tumor size, expression levels of Lewis y antigen and integrin α5β1 correlation with ovarian carcinoma chemotherapy resistance. Results: The expression rates of Lewis y antigen and integrins α5 and β1 were significantly greater in the drug-resistant group (91.17%, 85.29%, 88.24%) than the partially sensitive (50.00%, 33.33%, 50.00%) or sensitive groups (61.54%, 57.69%, 55.77%). Binary logistic regression analysis revealed that surgical stage, residual tumor size, and expression of integrin α5 and Lewis y in ovarian carcinoma tissues were independent risk factors for chemotherapeutic drug resistance. Conclusions: Overexpression of Lewis y and integrin α5 are strong risk factors for chemotherapeutic drug resistance in ovarian carcinoma patients.

The anti-carbamylated protein (CarP) antibody is a novel biomarker that might help in the diagnosis of rheumatoid arthritis (RA). We aim to assess the diagnostic value of anti-CarP antibody for RA.We systematically searched PubMed, Embase, the Cochrane Library, Web of Science, and Scopus for studies published by December 15, 2015. Studies in any language that evaluated the utility of the anti-CarP antibody in the diagnosis of RA in which healthy donors or patients without arthritis or arthralgia served as controls were included. Two investigators independently evaluated studies for inclusion, assessed study quality and abstracted data. A bivariate mixed-effects model was used to summarize the diagnostic indexes from 7 eligible studies.The pooled sensitivity, specificity, and positive and negative likelihood ratios for anti-CarP antibody were 42% (95% CI, 38% to 45%), 96% (95% CI, 95% to 97%), 10.2 (95% CI, 7.5 to 13.9), and 0.61 (95% CI, 0.57 to 0.65), respectively. The summary diagnostic odds ratio was 17 (95% CI, 12 to 24), and the area under summary receiver operator characteristic curve was 80% (95% CI, 77% to 84%).Anti-CarP antibody has a moderate value in the diagnosis of RA with high specificity but relatively low sensitivity.

Objective: To investigate the effect of Lewis y overexpression on the expression of proliferation-related factors in ovarian cancer cells. Methods: mRNA levels of cyclins, CDKs, and CKIs were measured in cells before and after transfection with the α1,2-fucosyltransferase gene by real-time PCR, and protein levels of cyclins, CDKs and CKIs were determined in cells before and after gene transfection by Western blot. Results: Lewis y overexpression led to an increase in both mRNA and protein expression levels of cyclin A, cyclin D1 and cyclin E in ovarian cancer cells, decrease in both mRNA and protein expression levels of p16 and p21, and decrease of p27 at only the protein expression level without change in its mRNA level. There were no differences in proteins and the mRNA levels of CDK2, CDK4 and CDK6 before and after gene transfection. Anti-Lewis y antibody, ERK and PI3K pathway inhibitors PD98059 and LY294002 reduced the difference in cyclin and CKI expression caused by Lewis y overexpression. Conclusion: Lewis y regulates the expression of cell cycle-related factors through ERK/MAPK and PI3K/Akt signaling pathways to promote cell proliferation.

Epithelial carcinomas of the ovary exhibit the highest mortality rate among gynecologic malignancies. Studies found that the metabolism of glycolipids or carbohydrates is associated with acquirement of anticancer drug-resistance by cancer cells. This study was to characterize possible involvement of Lewis Y (LeY) antigen in the drug-resistance of cancer cells. We transfected the α1,2-fucosyltransferase gene into human ovarian carcinoma-derived RMG-1 cells and established RMG-1-hFUT cells with enhanced expression of LeY. We determined the effects of docetaxel on the survival of cells by MTT assaying and observed the apoptosis of cells in the presence of docetaxel by flow cytometric analysis and by transmission electron microscopy. Plasma membranes and intracellular granules in RMG-1-hFUT cells were stained with anti-LeY antibody, the intensity of the staining was higher than that in control cells. The RMG-1-hFUT cells exhibited higher resistance to docetaxel than the control cells with regard to the docetaxel concentration and time course. After treatment with 10 μg/mL docetaxel for 72 h, the control cells, but not RMG-1-hFUT, contained abundant positively stained cell debris due to disintegration of the cytoskeleton. On transmission electron microscopy, although the control cells treated with docetaxel as above showed the following morphology, i.e., absence of villi, cells shrunken in size, pyknosis, agglutinated chromatin and cell buds containing nuclei in the process of apoptosis, the RMG-1-hFUT cells showed only agglutinated chromatin and vacuoles in the cytoplasm. In summary, cells with enhanced expression of LeY were shown to acquire docetaxel-resistance, indicating the possible involvement of glycoconjugates in the drug-resistance.

Objective: To detect the expression and clinical significances of Lewis y antigen and integrin αv, β3 in epithelial ovarian tumors, and to explore the expression correlation between Lewis y antigen and integrin αv, β3. Methods: Immunohistochemical staining was performed in 95 cases of epithelial ovarian cancer, 37 cases of borderline tumors, 20 cases of benign tumors, and 20 cases of normal ovarian tissue, for the detection of Lewis y antigen and integrin αv, β3 expressions, and to analyze the relationship between Lewis y antigen and integrin, and the relationship between clinical and pathological parameters of ovarian cancer. In addition, immunofluorescence double labeling was utilized to detect the expression correlation between Lewis y antigen and integrin αv, β3 in ovarian cancer. Results: In epithelial ovarian tumors, the expression rate of Lewis y antigen was 81.05%, significantly higher than that of borderline (51.53%) (P < 0.05) and benign (25%) (P < 0.01) tumors, and normal ovarian tissues (0) (P < 0.01). The expression rate of integrin αv, β3 in malignant epithelial ovarian tumors was 78.95% and 82.11%, respectively, significantly higher than that of the borderline (45.94%, 40.54%) (both P < 0.05), benign group (10.00%, 15.00%) (both P < 0.01) and normal ovary group (5%, 15%) (both P < 0.01). Conclusions: Lewis y and integrins αv, β3 are relevant to pelvic and abdominal diffusion and metastasis of ovarian cancer cells, suggesting that these two molecules mediate a boosting function for tumor metastasis.

Lewis (y) antigen is a difucosylated oligosaccharide present on the plasma membrane, and its overexpression is frequently found in human cancers and has been shown to be associated with poor prognosis. Our previous studies have shown that Lewis (y) antigen plays a positive role in the process of invasion and metastasis of ovarian cancer cells. However, the mechanisms by which Lewis (y) antigen enhances the invasion and tumor metastasis are still unknown. In this study, we established a stable cell line constitutively expressing Lewis (y) antigen (RMG-1-hFUT) by transfecting the cDNA encoding part of the human α1,2-fucosyltransferase (α1,2-FUT) gene into the ovarian cancer cell line RMG-1, and investigated whether Lewis (y) antigen regulates the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9, and tissue inhibitors of metalloproteinases (TIMP-1) and TIMP-2. We found that RMG-1-hFUT cells exhibited higher invasive capacities than their control cells. In addition, expression of TIMP-1 and TIMP-2 was down-regulated and expression of MMP-2 and MMP-9 was up-regulated. Anti-Lewis (y) antigen antibody treatment significantly reversed the expression of TIMP-1, TIMP-2, MMP-2 and MMP-9. Taken together, we provide the first evidence that down-regulation of TIMP-1 and TIMP-2 and up-regulation of MMP-2 and MMP-9 represents one of the mechanisms by which Lewis (y) antigen promotes cell invasion.

Lewis y (LeY) antigen is a difucosylated oligosaccharide carried by glycoconjugates on the cell surface. Overexpression of LeY is frequently observed in epithelial-derived cancers and has been correlated to the pathological staging and prognosis. However, the effects of LeY on ovarian cancer are not yet clear. Previously, we transfected the ovarian cancer cell line RMG-I with the α1,2-fucosyltransferase gene to obtain stable transfectants, RMG-I-H, that highly express LeY. In the present study, we examined the proliferation, tumorigenesis, adhesion and invasion of the cell lines with treatment of LeY monoclonal antibody (mAb). Additionally, we examined the expression of TGF-β1, VEGF and b-FGF in xenograft tumors. The results showed that the proliferation and adhesion in vitro were significantly inhibited by treatment of RMG-I-H cells with LeY mAb. When subcutaneously inoculated in nude mice, RMG-I-H cells produced large tumors, while mock-transfected cells RMG-I-C and the parental cells RMG-I produced small tumors. Moreover, the tumor formation by RMG-I-H cells was inhibited by preincubating the cells with LeY mAb. Notably, the expression of TGF-β1, VEGF and b-FGF all increased in RMG-I-H cells. In conclusion, LeY plays an important role in promoting cell proliferation, tumorigenecity and adhesion, and these effects may be related to increased levels of growth factors. The LeY antibody shows potential application in the treatment of LeY-positive tumors.

Several studies have identified an association between Behçet's disease (BD) and mutations in the Mediterranean fever (MEFV) gene, which was originally linked to the autosomal recessive disease, Familial Mediterranean fever. However, no consensus has been reached. Here, a meta-analysis was conducted on published data to comprehensively evaluate this relationship.Literature searches were performed in Pubmed, Embase, the Web of Science, and HuGE Navigator databases, in order to identify studies pertaining to the association between MEFV mutations and BD. Two investigators independently extracted and evaluated the data from eligible studies. The association between MEFV mutations (M694V, M680I, and E148Q) and BD was estimated overall by the odds ratio (OR) and 95% confidence intervals (95% CI). Further analysis was conducted with STATA 12.0 software (Stata Corp.; College Station, TX).Eligible studies (n=8) included genotyping data obtained from 2538 BD patients and 2792 healthy controls. Of the three mutations, M694V (pooled OR: 2.60, 95% CI: 2.02-3.34) and M680I (pooled OR: 1.74, 95% CI: 1.23-2.46) were found to be associated with BD in the overall analysis. The third mutation, E148Q, however, was not found to be linked with BD (pooled OR: 1.26, 95% CI: 0.69-2.31). Subgroup analysis furthermore revealed that M694V and M680I were risk loci for BD specifically in Turkish patients.The meta-analysis confirmed that MEFV mutations M694V and M680I were associated with BD. Additional studies from other ethnic populations and functional experiments are necessary to determine the extent to which the MEFV gene underlies the development of BD.

A previous large-scale replication study validation of a genome wide association study (GWAS) identified IκB kinase β (IKBKB) single nucleotide polymorphisms (SNPs) as a risk factor associated with systemic lupus erythematosus (SLE) in a Chinese Han population. IKBKB SNPs were associated with polymerase β (POLB) SNPs and reduced POLB expression, and this was proposed to be an underlying cause of human SLE development. In the current case-control study, we evaluated IKBKB (rs12676482 and rs2272733) and POLB (rs3136717 and rs3136744) SNPs in 946 SLE patients and 961 healthy controls. We investigated the possible association of these four SNPs with SLE in a Chinese Han population using the polymerase chain reaction-ligation detection reaction (PCR-LDR) technique. The differences in the frequencies of the four SNP alleles and the genotypes and haplotypes of the POLB polymorphisms were statistically insignificant when the SLE patients were compared with the controls in the Chinese Han population enrolled in this study (all, p ˃ 0.05). Furthermore, no associations were detected using different genetic models (additive, dominant, and recessive; all, p ˃ 0.05). Our findings indicate that the IKBKB (rs12676482 and rs2272733) and POLB (rs3136717, rs3136744) SNPs confer no genetic predisposition to SLE risk in this Chinese Han population.

The co-inhibitory receptor Programmed Death-1 (PD-1) curtails immune responses and prevent autoimmunity, however, tumors exploit this pathway to escape from immune destruction. The co-stimulatory receptor OX40 is upregulated on T cells following activation and increases their clonal expansion, survival and cytokine production when engaged. Although antagonistic anti-PD-1 or agonistic anti-OX40 antibodies can promote the rejection of several murine tumors, some poorly immunogenic tumors were refractory to this treatment. In the present study, we evaluated the antitumor effects and mechanisms of combinatorial PD-1 blockade and OX40 triggering in a murine ID8 ovarian cancer model. Although individual anti-PD-1 or OX40 mAb treatment was ineffective in tumor protection against 10-day established ID8 tumor, combined anti-PD-1/OX40 mAb treatment markedly inhibited tumor outgrowth with 60% of mice tumor free 90 days after tumor inoculation. Tumor protection was associated with a systemic immune response with memory and antigen specificity and required CD4(+) cells and CD8(+) T cells. The anti-PD-1/OX40 mAb treatment increased CD4(+) and CD8(+) cells and decreased immunosuppressive CD4(+)FoxP3(+) regulatory T (Treg) cells and CD11b(+)Gr-1(+) myeloid suppressor cells (MDSC), giving rise to significantly higher ratios of both effector CD4(+) and CD8(+) cells to Treg and MDSC in peritoneal cavity; Quantitative RT-PCR data further demonstrated the induction of a local immunostimulatory milieu by anti-PD-1/OX40 mAb treatment. The splenic CD8(+) T cells from combined mAb treated mice produced high levels of IFN-γ upon tumor antigen stimulation and exhibited antigen-specific cytolytic activity. To our knowledge, this is the first study testing the antitumor effects of combined anti-PD-1/OX40 mAb in a murine ovarian cancer model, and our results provide a rationale for clinical trials evaluating ovarian cancer immunotherapy using this combination of mAb.

OBJECTIVES:To measure Lewis y antigen and CD44 antigen expression in epithelial ovarian carcinoma and to correlate the levels of these antigens with clinical response to chemotherapy. METHODS:The study cases included 34 cases of ovarian carcinoma with resistance to chemotherapeutic drugs, 6 partially drug-sensitive cases, and 52 drug-sensitive cases (92 total). RESULTS:The rates of expression of Lewis y antigen and CD44 antigen were significantly greater in the drug-resistant group than that in the partially-sensitive or sensitive groups. Surgical stage, residual tumor size and expression of CD44 and Lewis y antigen in ovarian carcinoma tissues were independent risk factors for chemotherapeutic drug resistance. CONCLUSIONS:Over-expression of Lewis y and CD44 antigen are strong risk factors for chemotherapeutic drug resistance in ovarian carcinoma patients.

BACKGROUND: While HPV infection is the main cause of cervical cancer, genetic susceptibility to HPV infection is not well understood. Tumor necrosis factor alpha (TNF-alpha), involved in the defense against HPV infection, plays an important role in cervical cancer progression and regression. The aim of this study was to investigate the relationship between the TNF-alpha rs1800629 polymorphism and risk of HPV infection or cervical cancer. METHODS: Three groups were involved in this study of Chinese women. Group 1 consisted of 285 high risk HPV positive cervical cancer patients, Group 2, 225 high risk HPV positive patients without cervical cancer, and Group 3, 318 HPV negative women with no cervical cancer. Blood samples were obtained from all patients and genotyped by PCR-RLFP. Fifty randomly selected samples were further sequenced. RESULTS: The allele and genotype distributions of the TNF-alpha rs1800629 polymorphism were not significantly different between each of the groups (P>0.05). There are no significant relationship between rs1800629 polymorphism and high risk HPV infection (OR = 0.649, 95% CI: 0.253-1.670, P = 0.371), cervical cancer (OR = 0.993, 95% CI: 0.376-2.618, P = 0.988), or cervical cancer with HPV infection (OR = 0.663, 95% CI: 0.250-1.758, P = 0.409). CONCLUSIONS: We demonstrated that there is no association between TNF rs1800629 polymorphism and the HPV infection, or cervical cancer with HPV infection.

Soon after its independence from the British in August 1947, Pakistan's socio-political culture became entangled into a monopoly of the right-wing with feudal lords, especially in the Punjab province. The pseudo right-wing political parties exploited Pakistan's religious foundations and established a cultural hegemony. Historically, Punjab had remained the epicentre of political bargains since its inception; therefore, it retained strong alliances for stable government in the centre. Somehow, dissident voices and views from any other political ideology were deemed to be anti-state or anti-religious. This paper analyses through Antonio Gramsci's Theory of Cultural Hegemony how the right wing manipulated the opinion of the masses to legitimize their rule, which ultimately led to the decline of socialism. This paper illustrates the underlying ultra-religious campaigns in the electoral manoeuvring of the right wing from 1977 to 1997, thereby gaining the consent of the people. This study also outlines the systematic othering and demonizing of the left wing in society by pseudo right-wing factions. [ABSTRACT FROM AUTHOR]