Hemolytic disease of the newborn due to anti-Jra from a Chinese mother with one novel and one classic heterozygous mutation.

OBJECTIVE: Investigation of a Jr(a-) family samples, identification of the mutant and assessment of the differences of Jr antigen density of the Jr(a-) family members, random adult and newborn individuals' RBCs. BACKGROUND: The anti-Jra antibody is generated when a Jr(a-) individual pregnant or transfused with Jr(a+) blood unit, which can lead to mild-to-moderate hemolytic disease of the foetus and newborn (HDFN) or hemolytic transfusion reaction (HTR). Several mutations had been identified. The anti-Jra caused HDFN is not rare in East Asia, but due to the lack of antibody and molecular background, it is likely to lead missed detection. METHODS AND MATERIALS: One G4P1 woman had been detected as IAT positive during prenatal examination. Suspected as anti-Jra after the laboratory serological testing, the maternal sample was further assessed by molecular analysis. The antigen density was detected by flow cytometry after reacting with anti-Jra serum in family members and the normal individuals. RESULTS: One novel frameshift mutation c.717delC and one previously identified mutation c.706C > T in ABCG2 was identified on proband. The infant haemoglobin(Hb) and bilirubin increased significantly after exchange transfusion and the severe HDFN was relieved. Flow cytometry results showed that the Jra antigens on adult RBCs were significantly less than those on the infant. CONCLUSION: The c.717delC mutation can lead to the shortening of protein ABCG2 in the site of p.Leu307Stop, result in the loss of Jra antigen. The difference in antigen density between adult and infant RBCs may be a possible reason that leads to severe HDFN but not transfusion reaction. Breastfeeding may lead to slower recovery from HDFN.

Joint effects and mechanisms of luteolin and kaempferol on toxigenic Microcystis growth-Comprehensive analysis on two isomers interaction in binary mixture.

Allelochemicals are widely accepted as promising algaecide to mitigate Microcystis-dominated cyanobacterial blooms (MCBs). Allelopathic algicidal effect of single luteolin or kaempferol against Microcystis had been confirmed, but their joint effect against Microcystis was unclear. This study comprehensively explored time-dependent joint effect and mechanisms of luteolin and kaempferol on Microcystis growth during 14 day-test. The 50%-inhibitory threshold of their mixture (IC50 mix) was verified as 4.872 and 5.211 mg/L at equitoxic ratio, and 5.167 and 4.487 mg/L at equivalent ratio, respectively, on day 8 and 14. Using toxicity unit, isobologram and predictive models, results revealed that luteolin and kaempferol at equivalent ratio interacted additively at lower, median and higher dosages, while at equitoxic ratio interacted additively at lower dosage but synergistically at median and higher dosages in Microcystis on day 8 and 14, implying that their equitoxic mixture posed better algicidal effect against Microcystis. Various dosages of equitoxic mixture concurrently decreased aqueous and total microcystins (MCs) contents along test. Thus, luteolin and kaempferol could be jointly applied as high-efficacy and eco-safe algaecide with declined MCs pollution risks. As mixture dosage elevated, more strongly weakened cellular MCs retention and inhibited cellular photosynthetic pigments content during late stage, as well as decreased aqueous MCs content long test, jointly explained increasing growth inhibition ratio with rising mixture dosage. Yet, cell damage was gradually repaired due to early stimulated antioxidant defense at each mixture dosage, thus cell damage might not be a major reason for inhibited growth under mixture stress. This study provided novel insights and guidance to coupled application of luteolin and kamepferol for mitigating MCBs and decreasing MCs pollution risks.

A Network-Pharmacology-Combined Integrated Pharmacokinetic Strategy to Investigate the Mechanism of Potential Liver Injury due to Polygonum multiflorum.

Polygonum multiflorum (PM) has been used as a tonic and anti-aging remedy for centuries in Asian countries. However, its application in the clinic has been hindered by its potential to cause liver injury and the lack of investigations into this mechanism. Here, we established a strategy using a network pharmacological technique combined with integrated pharmacokinetics to provide an applicable approach for addressing this issue. A fast and sensitive HPLC-QQQ-MS method was developed for the simultaneous quantification of five effective compounds (trans-2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucoside, emodin-8-O-ß-d-glucoside, physcion-8-O-ß-d-glucoside, aloe-emodin and emodin). The method was fully validated in terms of specificity, linearity, accuracy, precision, extraction recovery, matrix effects, and stability. The lower limits of quantification were 0.125-0.500 ng/mL. This well-validated method was successfully applied to an integrated pharmacokinetic study of PM extract in rats. The network pharmacological technique was used to evaluate the potential liver injury due to the five absorbed components. Through pathway enrichment analysis, it was found that potential liver injury is primarily associated with PI3K-Akt, MAPK, Rap1, and Ras signaling pathways. In brief, the combined strategy might be valuable in revealing the mechanism of potential liver injury due to PM.

Rapid identification of chemical constituents of Rhodiola crenulata using liquid chromatography-mass spectrometry pseudotargeted analysis.

Rhodiola crenulata (R. crenulata), is a famous traditional Chinese medicine, with observable effects such as anti-high-altitude illness and fatigue resistance. Nevertheless, comprehensive and systematic structural identification of its components remains a challenge. In this study, a pseudotargeted analytical method, involving key fragment filtering by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry and ultra-high performance liquid chromatography-linear ion trap-Orbitrap mass spectrometry, was developed for rapid detection and identification of the chemical constituents of R. crenulata. The process consists of three steps: (i) acquiring sufficient mass spectral data, (ii) constructing a key fragments schedule and discovering the substructures rapidly by pseudotargeted key fragment filtering, and (iii) further identification of the compound structures based on accurate masses, fragment ions, related literatures, and authentic standards. As a result, 104 compounds were identified and divided into five categories, among which three potentially new and 59 ones were reported for the first time in R. crenulata. These results indicated that the major types of components are flavanols and gallic acid derivatives, organic acids, alcohols and their glycosides, flavonoids and their glycosides. This study enhances the understanding of R. crenulata and provides a reference for rapid and comprehensive identification of constituents in other herbal medicines.

Time- and dose-dependent allelopathic effects and mechanisms of kaempferol on toxigenic Microcystis growth.

This study determined time-dependent IC50 and confirmed 3.5 mg/L as IC50 value for kaempferol inhibiting toxigenic Microcystis growth, based on which algicidal effects and mechanisms against toxigenic Microcystis exposed to various kaempferol doses (0.5-2 × IC50) were explored along 14 day-test. Results showed that growth inhibition ratio (GIR) almost elevated with increasing kaempferol dose, and at each dose GIR elevated firstly and fluctuated around 17.8%- > 40%, 53.6%-65.6% and 84.8%-89.3% at 1.75, 3.5 and 7 mg/L kaempferol during mid-late stage, respectively. With rising kaempferol dose, photosynthetic pigments contents (chlorophyll-a, phycobiliproteins), antioxidant response (superoxide dismutase and catalase (CAT) activities, glutathione (GSH) contents) and microcystins (MCs) production were almost increasingly stimulated as cellular protective responses during early-mid stage. However, these parameters (excluding CAT and GSH) were almost increasingly inhibited at late stage by prolonged stress and Microcystis cell was still more severely damaged as dose elevated along test, which could be reasons for increasing GIR with rising kamepferol dose. Persistent stimulation of CAT and GSH at each dose could alleviate cell damage until late stage, thus GIR no longer increased at late stage at each kaempferol dose. Moreover, fewer MCs release under kaempferol stress than control suggested kaempferol as eco-safe algaecide for migrating toxigenic Microcystis-dominated blooms (MCBs) and decreasing MCs risks. Compared with our previous data for luteolin inhibiting toxigenic Microcystis, this study supported formerly-proposed 'flavonoids structure - algicidal activity' relationship that the only OH-location difference between kaempferol and luteolin could affect algicidal activity and mechanisms against toxigenic Microcystis. Also, kaempferol and luteolin was revealed to exert additive effect on toxigenic Microcystis growth at equitoxic ratio. Our findings gave novel algicidal scenario of flavonoids and were greatly implicated in eco-friendly migrating toxigenic MCBs.

Dual-ligand functionalized carbon nanodots as green fluorescent nanosensors for cellular dual receptor-mediated targeted imaging.

The conjugation of ligands to nanoparticles as drug delivery systems that target specific cells is a promising approach for the delivery of therapeutic agents to tumor cells. Herein, we prepared green-emission fluorescent carbon nanodots (CNDs) by a facile hydrothermal method with d-(+)-glucosamine hydrochloride and l-aspartic acid as the precursors, then covalently conjugated with folate (FA), polyethyleneimine (PEI) and hyaluronic acid (HA) to develop dual ligand-decorated nanocarriers (FA-PEI-HA-CNDs) for the targeted imaging of cancer cells. FA-PEI-HA-CNDs integrated the excellent fluorescence property of CNDs, and can be used for the real-time and noninvasive location tracking of cancer cells. The cellular uptake study demonstrated that FA-PEI-HA-CNDs markedly improved the internalization efficiency in A-549 cells via folate/CD44 receptor-mediated endocytosis in comparison with that of the A549 cells pretreated with excess FA, HA, and FA and HA. Therefore, these dual folate/CD44 receptor-targeted CNDs (FA-PEI-HA-CNDs) show promising potential for cancer detection, drug delivery, and individualized treatment as performance platforms.

OK/basigin expression on red blood cells varies between blood donors and correlates with binding of recombinant Plasmodium falciparum reticulocyte-binding protein homolog 5.

BACKGROUND: Recently, basigin (BSG), which carries OK antigens on red blood cells (RBCs), was reported to be the receptor of the Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRh5). BSG-PfRh5 is the only essential receptor-ligand pair in P. falciparum invasion that is known to date. However, the kind of OK/BSG polymorphism involved in the selection pressure caused by P. falciparum malaria has not been determined. STUDY DESIGN AND METHODS: Blood samples were collected to detect the expression of OK/BSG. The coding region of PfRh5 was cloned and expressed. Enzyme-linked immunosorbent assay-based erythrocyte binding assay was used to measure the recombinant PfRh5 (rPfRh5) binding of RBCs with different OK/BSG expressions. Sequencing of the BSG gene and quantification of the BSG mRNA were performed for selected samples. The candidate microRNAs (miRNAs), which might target the BSG gene, were obtained by miRNA sequencing. Dual-Luciferase reporter assay and overexpression of identified miRNAs were performed in K562 cells. RESULTS: The rPfRh5 was successfully expressed and verified. The OK/BSG expression levels varied among blood donors and were strongly associated with rPfRh5 binding. No single-nucleotide polymorphism was related to the OK/BSG expression. A potential BSG regulator, miR-501-3p, was identified by miRNA sequencing and Dual-Luciferase assay, but was not proven to regulate the expression of BSG in K562 cells. CONCLUSION: Although the mechanism of OK/BSG expression and regulation on RBCs has not been fully clarified, our findings suggest that the OK/BSG expression levels on RBCs might be related to P. falciparum invasion. Moreover, posttranscriptional regulation might play a role in controlling the OK/BSG expression.

[Studies on constituents of Polygonum multiflorum extract in vivo and in vitro based on UPLC-Q-TOF-MS].

To explore the drug-induced constituents in vivo of Polygonum multiflorum extract (PM). This study was the first to study the drug-induced constituents in target organ liver. Agilent MassHunter qualitative analysis software and Metabolite ID software were applied for the analysis of retention time, exact relative molecular mass, primary and secondary mass spectrum information based on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and targeted-MS/MS. By comparison with literature and standards, a total of 5 prototypes and 6 metabolites were identified or tentatively elucidated from the liver samples. In addition, the drug-induced constituents in plasma and PM were also analyzed in this study and 8 prototypes and 19 metabolites were detected from the plasma samples, while 30 compounds were detected from the extract of PM. Emodin oxidative acetylation metabolites, hydroxyl methylation metabolites, carboxylation glucuronidation metabolites and ketone glucuronidation metabolites in this study were first reported. Through the comparative analysis between the in vivo and in vitro constituents of PM, the study preliminarily revealed the drug-induced constituents (prototypes and metabolites) in liver and clarified the transfer process and transmutation rules of constituents in PM, blood and liver, which would further deepen our understanding on constituents of PM in vivo.

The Anti-hyperlipidemia Effects of Raw Polygonum multiflorum Extract in Vivo.

Polygonum multiflorum is widely used in the prevention and treatment of hyperlipidemia in traditional Chinese Medicine. In this study, the effects and relevant mechanisms of lipid-regulation by raw Polygonum multiflorum (RPM) were investigated. The results indicated that the basal plasma lipids, such as low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), and triglycerides (TG), were significantly decreased in RPM treatment groups compared with the model group, especially in the RPM high dose group. The key enzymes involved in lipid metabolism, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC) in plasma were generally reduced after oral administration, which was consistent with the transcription levels of their target genes. In addition, the hepatotoxicity of RPM was investigated, and RPM showed slightly less liver injury than that induced by simvastatin. Histological analysis indicated that the fat vacuoles and steatosis in hepatocytes were relieved after oral administration of RPM extract at a high dose of 16.2 g/kg, which was more obvious than that induced by simvastatin. These results revealed that RPM exerted its lipid-lowering effect by regulating the expression of related genes, and performed better than simvastatin in the treatment of hyperlipidemia.

[Effect of Zhuangyao Jianshen Wan (ZYJCW) on P2X1 and P2X3 mRNA expressions in rats with diuresis caused by kidney deficiency].

To investigate the urination-reducing effect and mechanism of Zhuangyao Jianshen Wan (ZYJCW). In this study, SI rats were subcutaneously injected with 150 mg · kg(-1) dose of D-galactose to prepare the sub-acute aging model and randomly divided into the model group, the Suoquan Wan group (1.17 g · kg(-1) · d(-1)), and ZYJCW high, medium and low dose groups (2.39, 1.20, 0.60 g · kg(-1) · d(-1)) , with normal rats in the blank group. They were continuously administered with drugs for eight weeks. The metabolic cage method was adopted to measure the 24 h urine volume and 5 h water load urine volume in rats. The automatic biochemistry analyzer was adopted to detect urine concentrations of Na+, Cl-, K+. The ELISA method was used to determine serum aldosterone (ALD) and antidiuretic hormone (ADH). The changes in P2X1 and P2X3 mRNA expressions in bladder tissues of rats were detected by RT-PCR. According to the results, both ZYJCW high and medium dose groups showed significant down-regulations in 24 h urine volume and 5 h water load urine volume in (P <0.05, P <0.01), declines in Na+ and Cl- concentrations in urine (P <0.01), notable rises in plasma ALD and ADH contents (P <0.05, P <0.01) and remarkable down-regulations in the P2X1 and P2X3 mRNA expressions in bladder tissues (P <0.01). The ZYJCW low dose group revealed obvious reductions in Na+ and Cl- concentrations in urine (P <0.01). The results indicated that ZYJCW may show the urination-reducing effect by down-regulating the P2X1 and P2X3 mRNA expressions in bladder tissues of rats with diuresis caused by kidney deficiency.

[Screening of rare blood group Lu(a-b-) phenotype and study of its molecular basis in ethnic Han Chinese from Shanghai region].

OBJECTIVE: To study the frequency of rare blood group Lu(a-b-) phenotype in a population from Shanghai region, and to explore the molecular basis of Lu(a-b-) by detecting the Lu and Lu relative mediator gene EKLF/KLF1. METHODS: Donors from Shanghai region were screened for Lutheran blood group by monoclonal anti-Lub using serological methods. Individuals with Lu(b-) were determined Lua, P1 and i antigens. Fifteen exons of the LU gene and 3 exons of the EKLF/KLF1 gene for the identified Lu(a-b-) samples were amplified and sequenced. RESULTS: Ten Lu(a-b-) donors were obtained from 44 331 donors from Shanghai region. No homozygous or heterozygous mutations were found in the LU gene, whilst 7 mutations in EKLF/KLF1 gene were identified in the 10 samples. CONCLUSION: The frequency of rare Lu(a-b-) blood group in Shanghai was approximately 0.02%, and all the individuals had an In(Lu) phenotype. The molecular basis of such samples may be related to mutations in the EKLF/KLF1 gene.

Modifying luteolin's algicidal effect on Microcystis by virgin and diversely-aged polystyrene microplastics: Unveiling novel mechanisms through microalgal adaptive strategies.

Luteolin has shown great potential in inhibiting Microcystis-dominated cyanobacterial blooms. However, widespread microplastics (MPs) in natural aquatic systems often serve as substrates for cyanobacterial growth, which could impact cyanobacterial resistance to external stresses and interfere with luteolin's algicidal effect. This study explored the influence of virgin and diversely-aged polystyrene microplastics (PS-MPs) on inhibitory effect of luteolin on Microcystis growth and its microcystins (MCs) production/release. Moreover, the underlying mechanisms were also revealed by jointly analyzing SEM image, antioxidant response, exopolymeric substances (EPSs) production, and functional gene expression. Results suggested that 0.5, 5, and 50 mg/L virgin and diversely-aged PS-MPs almost weakened growth inhibition and oxidative damage of two doses of luteolin against Microcystisby stimulating its EPSs production and inducing self-aggregation of Microcystis cells and/or hetero-aggregation between Microcystis cells and PS-MPs. Compared to virgin PS-MPs, photo-aged PS-MPs possessed rougher flaky surfaces, and hydrothermal-aged PS-MPs showed internal cracking. These characteristics led to greater stimulation of EPS production and exhibited more significant protective effects on Microcystis. Notably, PS-MPs also decreased MCs content in aqueous phase, likely because they adsorbed some MCs. Such toxigenic hetero-aggregates formed by MCs, MPs, and Microcystis cells would directly poison grazing organisms that consume them and create more pathways for MCs into food web, posing greater eco-risks. This is the first study to clarify the influence and mechanisms of virgin and diversely-aged MPs on allelopathic algicidal effects from the perspective of microalgal inherent adaptive strategies.

Meta-analysis for systematic review of global micro/nano-plastics contamination versus various freshwater microalgae: Toxicological effect patterns, taxon-specific response, and potential eco-risks.

Micro/nano-plastics (MNPs), as emerging persistent pollutants, are threatening freshwater ecosystems worldwide. Microalgae are important primary producers at the base of trophic level and susceptible to MNPs contamination, possibly resulting in further contamination in higher trophic levels and water quality. This study conducted a systematic review of 1071 observations from 63 publications, utilizing meta-analysis and subgroup analysis to investigate the toxicological effect patterns of MNPs parameters (size, concentration, and type) on microalgae. We also explored the potential eco-risks of certain specific MNPs parameters and subtle variations in the response of various microalgae taxa to MNPs. Results suggested that microplastics significantly inhibited microalgal photosynthesis, while nano-plastics induced more severe cell membrane damage and promoted toxin-release. Within a certain range of concentrations (0∼50 mg/L), rising MNPs concentration progressively inhibited microalgal growth and chlorophyll-a content, and progressively enhanced toxin-release. Among MNPs types, polyamide caused higher growth inhibition and more severe lipid peroxidation, and polystyrene induced more toxin-release, whereas polyethylene terephthalate and polymethyl methacrylate posed minimal effects on microalgae. Moreover, Bacillariophyta growth was inhibited most significantly, while Chlorophyta displayed strong tolerance and Cyanophyta possessed strong adaptive and exceptional resilience. Particularly, Komvophoron, Microcystis, Nostoc, Scenedesmus, and Gomphonema were more tolerant and might dominate freshwater microalgal communities under MNPs contamination. These results are crucial for acquiring the fate of freshwater microalgae under various MNPs contamination, identifying dominant microalgae, and reasonably assessing and managing involved eco-risks.

Identifying rice field weeds from unmanned aerial vehicle remote sensing imagery using deep learning.

BACKGROUND: Rice field weed object detection can provide key information on weed species and locations for precise spraying, which is of great significance in actual agricultural production. However, facing the complex and changing real farm environments, traditional object detection methods still have difficulties in identifying small-sized, occluded and densely distributed weed instances. To address these problems, this paper proposes a multi-scale feature enhanced DETR network, named RMS-DETR. By adding multi-scale feature extraction branches on top of DETR, this model fully utilizes the information from different semantic feature layers to improve recognition capability for rice field weeds in real-world scenarios. METHODS: Introducing multi-scale feature layers on the basis of the DETR model, we conduct a differentiated design for different semantic feature layers. The high-level semantic feature layer adopts Transformer structure to extract contextual information between barnyard grass and rice plants. The low-level semantic feature layer uses CNN structure to extract local detail features of barnyard grass. Introducing multi-scale feature layers inevitably leads to increased model computation, thus lowering model inference speed. Therefore, we employ a new type of Pconv (Partial convolution) to replace traditional standard convolutions in the model. RESULTS: Compared to the original DETR model, our proposed RMS-DETR model achieved an average recognition accuracy improvement of 3.6% and 4.4% on our constructed rice field weeds dataset and the DOTA public dataset, respectively. The average recognition accuracies reached 0.792 and 0.851, respectively. The RMS-DETR model size is 40.8 M with inference time of 0.0081 s. Compared with three classical DETR models (Deformable DETR, Anchor DETR and DAB-DETR), the RMS-DETR model respectively improved average precision by 2.1%, 4.9% and 2.4%. DISCUSSION: This model is capable of accurately identifying rice field weeds in complex real-world scenarios, thus providing key technical support for precision spraying and management of variable-rate spraying systems.

Correlation Analysis of IL-17, IL-21, IL-23 with Non-Alcoholic Liver Fibrosis and Cirrhosis.

Objective: This research aimed to explore the involvement of interleukins (IL) - IL-6, IL-17, IL-21, and IL-23 - in the evolution and diagnosis of non-alcoholic liver fibrosis and cirrhosis. Methods: The study subjects were selected from the patients who visited the Department of Hepatology of X Hospital in Y City from August 2021 to April 2023. Peripheral blood samples were collected. All participants were divided into liver fibrosis, cirrhosis, hepatitis, and healthy subjects four groups. IL-21, IL-17, IL23, IL-6 were detected by double antibody sandwich. Results: The results showed that there was a significant difference in the levels of IL-17, IL-21, and IL-23 among the 4 groups (P<0.0001). ROC curve analysis showed that the AUC values of IL-17, IL-21 and liver fiber 4 items were >0.70, suggesting that the diagnostic efficacy of IL-17, IL-21 was similar to that of liver fiber 4 items. Spearman correlation analysis showed that IL-17 had a positive correlation with collagen type III N-peptide, type IV collagen, and Laminin (P < 0.05), and no correlation with Hyaluronic acid (P > 0.05). Conclusion: IL-17, IL-21, and IL-23 play a pivotal role in the inflammatory pathways associated with liver injuries, establishing themselves as potent auxiliary diagnostic markers in identifying liver fibrosis and cirrhosis.

Discovery of 2,8-dihydroxyadenine in HUA patients with uroliths and biomarkers for its associated nephropathy.

Currently, it is acknowledged that gout is caused by uric acid (UA). However, some studies have revealed no correlation between gout and UA levels, and growing evidence suggests that 2,8-dihydroxyadenine (2,8-DHA), whose structural formula is similar to UA but is less soluble, may induce gout. Hence, we hypothesized that uroliths from hyperuricemia (HUA) patients, which is closely associated with gout, may contain 2,8-DHA. In this study, 2,8-DHA in uroliths and serum of HUA patients were determined using HPLC. Moreover, bioinformatics was used to investigate the pathogenic mechanisms of 2,8-DHA nephropathy. Subsequently, a mouse model of 2,8-DHA nephropathy established by the gavage administration of adenine, as well as a model of injured HK-2 cells induced by 2,8-DHA were used to explore the pathogenesis of 2,8-DHA nephropathy. Interestingly, 2,8-DHA could readily deposit in the cortex of the renal tubules, and was found in the majority of these HUA patients. Additionally, the differentially expressed genes between 2,8-DHA nephropathy mice and control mice were found to be involved in inflammatory reactions. Importantly, CCL2 and IL-1ß genes had the maximum degree, closeness, and betweenness centrality scores. The expressions of CCL2 and IL-1ß genes were significantly increased in the serum of 24 HUA patients with uroliths, indicating that they may be significant factors for 2,8-DHA nephropathy. Further analysis illustrated that oxidative damage and inflammation were the crucial processes of 2,8-DHA renal injury, and CCL2 and IL-1ß genes were verified to be essential biomarkers for 2,8-DHA nephropathy. These findings revealed further insights into 2,8-DHA nephropathy, and provided new ideas for its diagnosis and treatment.

Weak D phenotypes caused by intronic mutations in the RHD gene: four novel weak D alleles identified in the Chinese population.

BACKGROUND: Although more than 80 weak D types have been reported, many rare alleles probably remain unidentified. However, direct evidence that associates intronic mutations in the RHD gene with weak D types is lacking. STUDY DESIGN AND METHODS: Blood samples were obtained from Shanghai Blood Center. D- samples typed in routine laboratories were tested using a monoclonal immunoglobulin M reagent, an indirect antiglobulin test, and a commercial panel of monoclonal anti-D (Diagast D-Screen). RHD nucleotide sequencing that included adjacent flanking intron regions was performed. The RHD zygosity was determined. Bioinformatics analysis was performed. RESULTS: Four different RHD alleles were identified among six blood samples, namely, RHD IVS3+3G>C, RHD IVS6-14del3, RHD IVS4+5G>A, and RHD IVS4+5G>T. The serologic tests were consistent with weak D phenotypes. The bioinformatics results indirectly suggest that these sequence changes affect splicing. CONCLUSION: This study is the first to describe weak D types caused by intronic variations near the splice sites in the RHD gene, which is supported by the genotyping results combined with serologic profiles and bioinformatics analysis. The identification of the four novel RHD alleles represents a new significant addition to the molecular bases that underlie weak D, which would deepen our understanding of the mechanism of the low expression of the RhD antigen. These nucleotide changes are predicted to have regulatory effects on RHD splicing.

The mutation spectrum of the JK-null phenotype in the Chinese population.

BACKGROUND: This study aimed to analyze the mutation spectrum of the JK-null phenotype in the Chinese population. The JK gene encoding the Kidd blood group antigen protein and JK*A/JK*B polymorphism caused by a G-to-A mutation at nt838 are well described. However, the molecular basis of the JK-null phenotype in Chinese populations remains unclear. STUDY DESIGN AND METHODS: Sixteen unrelated JK-null phenotype donors detected by red blood cell urea lysis resistance assay of 201,194 Chinese blood donors were confirmed in serologic agglutination tests. JK-null alleles were analyzed by MnlI polymerase chain reaction-restriction fragment length polymorphism and sequencing of all JK gene coding regions. RESULTS: In addition to the well-known Polynesian JK-null allele JK*B(IVS5-1g>a) and two alleles discovered in Taiwan, JK*B(896G>A) and JK*B(222C>A), seven JK-null allele types were detected in this study including four novel JK-null alleles: a nonsense mutation, JK*B(512G>A); two types of missense point mutations, JK*B(536C>G) and JK*B(437T>C); and a splice mutation, JK*A(IVS8+5g>a), resulting in skipping of Exon 8. CONCLUSION: This study demonstrates the frequency and heterogeneity of the JK-null phenotype in Chinese populations. Based on our findings, the mechanisms underlying the Chinese Jk(a-b-) phenotype are quite different from other ethnic groups. The two most common types of JK-null alleles were JK*B(IVS5-1g>a) and JK*B(896G>A) in Chinese persons. Four novel JK-null alleles were noted to be associated with the Jk(a-b-) phenotype.

Novel ecological implications of non-toxic Microcystis towards toxic ecotype in population-promoting toxic ecotype dominance at various N levels and cooperative defense against luteolin-stress.

Microcystin (MC)-producing (MC+) and MC-free (MC-) Microcystis always co-exist and interact during Microcystis-dominated cyanobacterial blooms (MCBs), where MC+Microcystis abundance and extracellular MC-content (EMC) determine the hazard extent of MCBs. The current study elucidated intraspecific interaction between MC+ and MC-Microcystis at various nitrogen (N) levels (0.5-50 mg/L) and how such N-mediated interaction impacted algicidal and EMC-inhibiting effect of luteolin, a natural bioalgicide. Conclusively, MC+ and MC-Microcystis were inhibited mutually at N-limitation (0.5 mg/L), which enhanced the algicidal and EMC-inhibiting effects of luteolin. However, at N-sufficiency (5-50 mg/L), MC-Microcystis promoted MC+ ecotype growth and dominance, and such intraspecific interaction induced the cooperative defense of two ecotypes, weakening luteolin's algicidal and EMC-inhibiting effects. Mechanism analyses further revealed that MC+Microcystis in luteolin-stress co-culture secreted exopolymeric substances (EPSs) for self-protection against luteolin-stress and also released more EMC to induce EPS-production by MC-Microcystis as protectants, thus enhancing their luteolin-resistance and promoting their growth. This study provided novel ecological implications of MC-Microcystis toward MC+ ecotype in terms of assisting the dominant establishment of MC+Microcystis and cooperative defense with MC+ ecotype against luteolin, which guided the application of bioalgicide (i.e. luteolin) for MCBs and MCs pollution mitigation in different eutrophication-degree waters.

Long-term tracking robust removal of Microcystis-dominated bloom and microcystin-pollution risk by luteolin continuous-release microsphere at different nitrogen levels-Mechanisms from proteomics and gene expression.

Luteolin continuous-release microsphere (CRM) has promising algicidal effect against Microcystis, but how nitrogen (N) level impacted CRM effects on Microcystis growth and microcystins (MCs) pollution was never tracked along long term. This study revealed that luteolin CRM exerted long-term and robust inhibitory effects on Microcystis growth and MC-pollution by sharply decreasing extracellular and total MCs content at each N level, with growth inhibition ratio of 88.18%-96.03%, 92.91%-97.17% and 91.36%-95.55% at 0.5, 5 and 50 mg/L N, respectively, during day 8-30. Further analyses revealed that CRM-stress inhibited transferase, GTPase and ATPase activities, ATP binding, metal ion binding, fatty acid biosynthesis, transmembrane transport and disrupted redox homeostasis to pose equally robust algicidal effect at each N level. At lower N level, CRM-stress tended to induce cellular metabolic mode towards stronger energy supply/acquisition but weaker energy production/consumption, while triggered a shift towards stronger energy production/storage but weaker energy acquisition/consumption as N level elevated, thus disturbing metabolic balance and strongly inhibiting Microcystis growth at each N level. Long-term robust algicidal effect of CRM against other common cyanobacteria besides Microcystis was evident in natural water. This study shed novel insights into inhibitory effects and mechanisms of luteolin CRM on Microcystis growth and MC-pollution in different N-level waters.