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Abstract Background Polycystic ovarian syndrome (PCOS) accounts for about 75% of anovulatory infertility. The cause of PCOS is not clear. CircRNAs acting as miRNA sponges mediate the post-transcriptional regulation of multiple genes. CYP19A1 is a limiting enzyme in the ovarian steroidogenesis pathway. However, the mechanism of circRNAs regulating granulosa cell (GC) estradiol secretion in PCOS remains to be elucidated. Methods Bioinformatics was used to predict the potential target miRNAs of circ_0043532 and target genes of miR-1270. Target miRNAs and mRNA expression were verified by qRT-PCR in GCs from 45 women with PCOS and 65 non-PCOS. Western blot, ELISA and dual-luciferase reporter assays were applied to confirm the substrate of miR-1270. Results Circ_0043532 and CYP19A1 were significant up-regulation in GCs from patients with PCOS. The predicted target miRNAs of circ_0053432, miR-1270, miR-576-5p, miR-421 and miR-142-5p, were notably decreased in GCs from patients with PCOS. Mechanistic experiments showed that circ_0043532 specifically binds to miR-1270. MiR-1270 was negatively regulated by circ_0043532. Concomitantly, miR-1270 inhibited CYP19A1 expression and estradiol production, which could be reversed by circ_0043532 over-expression. Conclusion We identified that circ_0043532/miR-1270/CYP19A1 axis contributes to the aberrant steroidogenesis of GCs from patients with PCOS. This study broadens the spectrum of pathogenic factors of PCOS, and circ_0043532 might be a potential therapeutic target for PCOS.

Abstract Background Methylmalonic aciduria (MMA) is a group of rare genetic metabolic disorders resulting from defects in methylmalonyl coenzyme A mutase (MCM) or intracellular cobalamin (cbl) metabolism. MMA patients show diverse clinical and genetic features across different subtypes and populations. Methods We retrospectively recruited 60 MMA patients from a single center and diagnosed them based on their clinical manifestations and biochemical assays. We then performed genetic analysis to confirm the diagnosis and identify the causal variants. Results We confirmed the common clinical manifestations of MMA reported previously. We also described four rare MMA cases with unusual symptoms or genetic variants, such as pulmonary hypertension or limb weakness in late-onset patients. We identified 15 MMACHC and 26 MMUT variants in 57 patients, including 6 novel MMUT variants. Two patients had only one MMAA variant each, and one patient had mild MMA due to mitochondrial DNA depletion syndrome caused by a SUCLA2 variant. Among 12 critically ill patients, isolated MMA was associated with higher C3, blood ammonia, and acidosis, while combined MMA was linked to hydrocephalus on skull MRI. MMACHC c.658-660delAAG and MMUT c.1280G > A variants were correlated with more severe phenotypes. Conclusions Our study demonstrates the clinical and genotypic heterogeneity of MMA patients and indicates that metabolic screening and genetic analysis are useful tools to identify rare cases.

ABSTRACT African swine fever (ASF), caused by ASF virus (ASFV), is a highly infectious and severe hemorrhagic disease of pigs that causes major economic losses. Currently, no commercial vaccine is available and prevention and control of ASF relies mainly on early diagnosis. Here, a novel automated double antigen sandwich chemiluminescent immunoassay (DAgS-aCLIA) was developed to detect antibodies against ASFV p72 (p72-Ab). For this purpose, recombinant p72 trimer was produced, coupled to magnetic particles as carriers and labeled with acridinium ester as a signal trace. Finally, p72-Ab can be sensitively and rapidly measured on an automated chemiluminescent instrument. For quantitative analysis, a calibration curve was established with a laudable linearity range of 0.21 to 212.0 ng/mL (R2 = 0.9910) and a lower detection limit of 0.15 ng/mL. For qualitative analysis, a cut-off value was set at 1.50 ng/mL with a diagnostic sensitivity of 100.00% and specificity of 98.33%. Furthermore, antibody response to an ASF gene-deleted vaccine candidate can be accurately quantified using this DAgS-aCLIA, as evidenced by early seroconversion as early as 7 days post-immunization and high antibody levels. Compared with available enzyme-linked immunosorbent assays, this DAgS-aCLIA demonstrated a wider linearity range of 4 to 16-fold, and excellent analytical sensitivity and agreement of over 95.60%. In conclusion, our proposed DAgS-aCLIA would be an effective tool to support ASF epidemiological surveillance.IMPORTANCEAfrican swine fever virus (ASFV) is highly contagious in wild boar and domestic pigs. There is currently no vaccine available for ASF, so serological testing is an important diagnostic tool. Traditional enzyme-linked immunosorbent assays provide only qualitative results and are time and resource consuming. This study will develop an automated chemiluminescent immunoassay (CLIA) that can quantitatively and qualitatively detect antibodies to ASFV p72, greatly reducing detection time and labour-intensive operation, and improving detection sensitivity and linearity range. This novel CLIA would serve as a reliable and convenient tool for ASF pandemic surveillance and vaccine development.

Water processes secure plant survival and maintain their ecosystem function. Salinity affects water processes, but the mechanisms remain unclear and may depend on the degree of salinity stress. To improve the understanding of the cooperation of plant organs involved in water processes under salinity stress, we determined hydraulic, gas exchange, and physiological and biochemical parameters in Populus euphratica Oliv. under different salinity stresses. The results suggested that P. euphratica enhanced water transport efficiency in a salinity-stress environment, and the strengthening effect of roots in the water transfer process was greater than that of the aboveground parts. P. euphratica also increased water use efficiency and water transport efficiency in mild and moderate salinity stress (less than 200 mmol/L NaCl) but was adversely affected by heavy salinity stress (more than 300 mmol/L NaCl). Furthermore, P. euphratica increased its water storage by regulating antioxidant enzyme scavenging capacity and osmoregulation, which resulted in coordinated greater water utilization and enhanced water transport among plant organs and indicated that the adverse effects on water processes triggered by salinity stress depended on the extent of salt stress. P. euphratica lessened stress-induced damage and maintained plant productivity by coordination and cooperation of water processes under certain levels of salinity. Research on the coordination and cooperation involving water processes in riparian forests in saline areas provides the scientific basis for riparian plant protection and restoration.

IntroductionIntrauterine transfusion of platelet-rich plasma (PRP) has become a new treatment for thin endometrium (TE) in recent years, but its low efficacy due to rapid release of growth factors limits its clinical use. Platelet-rich fibrin (PRF) starts the coagulation cascade reaction immediately after the blood comes into contact with the test tube. The natural coagulation process results in stable platelet activation and the slow release of growth factors.MethodsIn our study, primary human endometrial stromal cells (hESCs) were extracted from endometrial tissue. PRP and PRF were prepared from the patient cubital vein blood. Stromal cells were cultured in conditioned medium supplemented with PRP and PRF. Differences in cell behavior were observed by cell proliferation test and cell migration test. The relative expression levels of apoptotic Bax and antiapoptotic Bcl-2 genes were measured by qRT-PCR. The release of growth factors from PRP and PRF was detected by ELISA.ResultsWe found that both PRP and PRF inhibited apoptosis of hESCs, which favored cell proliferation and migration. In addition, PRF releases growth factors for a longer period of time compared to PRP.DiscussionPRF offer a more sustained therapeutic effect compared to PRP, which provides a new idea for endometrial regeneration and repair.

Abstract Background Recent studies have more focused on gut microbial alteration in tuberculosis (TB) patients. However, no detailed study on gut fungi modification has been reported till now. So, current research explores the characteristics of gut microbiota (bacteria)- and mycobiota (fungi)-dysbiosis in TB patients and also assesses the correlation between the gut microbiome and serum cytokines. It may help to screen the potential diagnostic biomarker for TB. Results The results show that the alpha diversity of the gut microbiome (including bacteria and fungi) decreased and altered the gut microbiome composition of TB patients. The bacterial genera Bacteroides and Prevotella were significantly increased, and Blautia and Bifidobacterium decreased in the TB patients group. The fungi genus Saccharomyces was increased while decreased levels of Aspergillus in TB patients. It indicates that gut microbial equilibrium between bacteria and fungi has been altered in TB patients. The fungal-to-bacterial species ratio was significantly decreased, and the bacterial-fungal trans-kingdom interactions have been reduced in TB patients. A set model including Bacteroides, Blautia, Eubacterium_hallii_group, Apiotrichum, Penicillium, and Saccharomyces may provide a better TB diagnostics option than using single bacterial or fungi sets. Also, gut microbial dysbiosis has a strong correlation with the alteration of IL-17 and IFN-γ. Conclusions Our results demonstrate that TB patients exhibit the gut bacterial and fungal dysbiosis. In the clinics, some gut microbes may be considered as potential biomarkers for auxiliary TB diagnosis.

相较于甘油三酯脂肪酶，偏甘油酯脂肪酶因其独特的底物选择性和催化高效性，逐渐成为相关领域的研究热点。旨在为偏甘油酯脂肪酶在油脂加工、脂质分子的定向合成等领域的研究提供理论和技术参考，并为挖掘新型的偏甘油酯脂肪酶打下基础，首先对偏甘油酯脂肪酶的来源和分类进行介绍，随后阐述了偏甘油酯脂肪酶的结构特征和催化特性，最后对其应用现状进行了总结归纳和展望。偏甘油酯脂肪酶有动物来源、植物来源和微生物来源，分为甘油单酯脂肪酶和甘油单酯-甘油二酯脂肪酶；偏甘油酯脂肪酶有帽区、催化三联体和氧负离子洞穴等结构，具有底物特异性和位置特异性；偏甘油酯脂肪酶在食品、有机合成、医疗等领域有着广泛的应用和独特的发展潜力。Compared with triglyceride lipase, partial glyceride lipase has gradually become a hot research focus in related fields due to its unique substrate selectivity and high catalytic efficiency. In order to provide theoretical and technical references for the research of partial glyceride lipase in the fields of oil processing and directed synthesis of lipids, and lay a foundation for excavating new partial glyceride lipase, firstly, the source and classification of partial glyceride lipase were introduced; subsequently, the structural and catalytic characteristics of partial glyceride lipase were expounded; finally its application status was summarized and prospected. Partial glyceride lipase comes from animal, plant and microbe, and is divided into monoacyglyceride lipase and glycerol mono-and diglyceride lipase. Partial glyceride lipase has structures of a cap region, catalytic triplet, and oxygen anion cave, and it has substrate specificity and site specificity. Partial glyceride lipase has extensive applications and unique development potential in fields such as food, organic synthesis, and medicine.