Analysis of risk factors for carotid artery plaque in asymptomatic adults.

OBJECTIVE: This study aimed to investigate the risk factors affecting the presence of carotid plaque in asymptomatic adults. METHODS: Asymptomatic adults (age > 40 years, no symptoms of cardiovascular and cerebrovascular diseases) undergoing routine health examinations from physical examination department were included in this study. Carotid plaque was measured by Resona 7OB and Resona 8EXP color Doppler ultrasound and L9-3U and L4-5WU probes. The focal carotid intima-media thickness was greater than 1.1 mm, and the local protrusion of the artery wall into the artery lumen suggested the presence of carotid atherosclerotic plaque. According to their ultrasound results, 1077 asymptomatic adults were divided into a group with carotid plaque (477) and a group without carotid plaque (600). RESULTS: A total of 1077 asymptomatic adults were included in this study, of whom 44.3% had carotid plaque. The proportion of men with carotid plaque was 84.5%. Multifactorial logistic analysis suggested that age, fasting blood glucose (FBG), total cholesterol (TC), homocysteine (Hcy) and male gender were risk factors for carotid atherosclerosis. The predictive probability of these risk factor indicators derived from the multifactorial model was calculated using receiver operating characteristic (ROC) curves with SPSS 25.0 software. The calculated area under the receiver operating characteristic curve (AUC) was 0.715 (95% CI, 0.685-0.746). CONCLUSION: Age, FBG, TC, Hcy and male gender are risk factors for carotid atherosclerosis in asymptomatic adults. Gender differences in carotid atherosclerosis deserve further attention.

Prognostic outcome of intravenous thrombolysis in elderly patients aged ≥ 60 years with acute ischemic stroke by ASTRAL and THRIVE scales.

This study aimed to validate the predictive performance of ASTRAL and THRIVE scales when used for patients aged 60 years and older with acute ischemic stroke (AIS) after intravenous thrombolysis (IVT). All enrolled patients received IVT therapy. The enrolled patients were divided into two groups in accordance with the modified Rankin scale(mRS) score at the time of discharge: good-outcome (mRS ≤ 2) and poor-outcome (mRS ≥ 3) groups. The receiver operating characteristic (ROC) curve was plotted using MedCalc software, the area under the ROC curve (AUC) was calculated. The Delong test was used to compare the predictive performance of ASTRAL and THRIVE scales, with P < 0.05 being considered a statistically significant difference. The AUCs of ASTRAL and THRIVE in predicting poor outcomes after thrombolysis in elderly patients with AIS were 0.771 and 0.701, respectively. The difference in AUC between ASTRAL and THRIVE was 0.070, and a statistically significant difference (P < 0.05) was found. ASTRAL's predictive performance was better than that of THRIVE. ASTRAL is a reliable predictive tool for assessing the poor outcome of IVT therapy for elderly patients aged ≥ 60 years with AIS.

ASTRAL scale for predicting prognosis following intravenous thrombolysis with anterior versus posterior circulation acute ischemic stroke.

In this study, we compared whether there was any difference between the ASTRAL(Acute Stroke Registry and Analysis of Lausanne, ASTRAL) scale in predicting prognosis after IVT(Intravenous Thrombolysis, IVT) in patients with AIS(Acute Ischemic Stroke, AIS) in the ACI(Anterior Circulation Infarction, ACI) and PCI(Posterior Circulation Infarction, PCI), with the aim of providing more guiding information. Statistical analysis was performed using SPSS 25.0. When comparing the baseline characteristics, the normal distribution test was carried out first, which did not conform to the normal distribution. The continuous variables were expressed in the median and interquartile, and the nonparametric double-independent sample test was carried out. MedCalc software was used to plot ROC(Receiver Operating Characteristic, ROC) curves, calculate AUC(Area Under the Receiver Operating Characteristic Curve, AUC), and compare the prediction performance of the ASTRAL score by Delong text, and the difference of P < 0.05 was statistically significant. The AUCs of ASTRAL in predicting poor prognosis of ACI and PCI patients after IVT were 0.768 and 0.773, respectively. There was no difference in the AUC of ASTRAL score between ACI and PCI(P > 0.05). The ASTRAL scale has consistent prognostic predictive value for AIS in the anterior and posterior circulatory systems and is a reliable tool for predicting poor prognosis of patients with ACI and PCI after IVT.

DhFIG_2, a gene of nematode-trapping fungus Dactylellina haptotyla that encodes a component of the low-affinity calcium uptake system, is required for conidiation and knob-trap formation.

Calcium ion (Ca2+) is a universal second messenger involved in regulating diverse processes in animals, plants, and fungi. The low-affinity calcium uptake system (LACS) participates in acquiring Ca2+ from extracellular environments under high extracellular Ca2+ concentration. Unlike most fungi, which encode only one protein (FIG1) for LACS, nematode-trapping fungi (NTF) encode two related proteins. AoFIG_2, the NTF-specific LACS component encoded by adhesive network-trap forming Arthrobotrys oligospora, was shown to be required for conidiation and trap formation. We characterized the role of DhFIG_2, an AoFIG_2 ortholog encoded by knob-trap forming Dactylellina haptotyla, in growth and development to expand our understanding of the role of LACS in NTF. Because repeated attempts to disrupt DhFIG_2 failed, knocking down the expression of DhFIG_2 via RNA interference (RNAi) was used to study its function. RNAi of DhFIG_2 significantly decreased its expression, severely reduced conidiation and trap formation, and affected vegetative growth and stress responses, suggesting that this component of LACS is crucial for trap formation and conidiation in NTF. Our study demonstrated the utility of RNAi assisted by ATMT for studying gene function in D. haptotyla.

DdaSTE12 is involved in trap formation, ring inflation, conidiation, and vegetative growth in the nematode-trapping fungus Drechslerella dactyloides.

Ste12 transcription factors, downstream of mitogen-activated protein kinase (MAPK) signalling pathways, are exclusively found in the fungal kingdom and regulate fungal mating, development, and pathogenicity. The nematode-trapping fungus Drechslerella dactyloides can capture free-living nematodes using constricting rings by cell inflation within 1 s when stimulated by nematodes entering the rings. The MAPK signalling pathways are involved in the trap formation of nematode-trapping fungi, but their downstream regulation is not clearly understood. In this study, disruption of the DdaSTE12 gene in D. dactyloides disabled cell inflation of constricting rings and led to an inability to capture nematodes. The number of septa of constricting rings and the ring cell vacuoles were changed in ΔDdaSTE12. Compared with the wild type, ΔDdaSTE12 reduced trap formation, conidiation, and vegetative growth by 79.3%, 80.3%, and 21.5%, respectively. The transcriptomes of ΔDdaSTE12-3, compared with those of the wild type, indicated that the expression of genes participating in trap formation processes, including signal transduction (Gpa2 and a 7-transmembrane receptor), vesicular transport and cell fusion (MARVEL domain-containing proteins), and nematode infection (PEX11 and CFEM domain-containing proteins), is regulated by DdaSTE12. The results suggest that DdaSTE12 is involved in trap formation and ring cell inflation, as well as conidiation and vegetative growth, by regulating a wide range of downstream functions. Our findings expanded the roles of Ste12 homologous transcription factors in the development of constricting rings and provided new insights into the downstream regulation of the MAPK signalling pathway involved in nematode predation. KEY POINTS: • DdaSTE12 was the first gene disrupted in D. dactyloides. • DdaSTE12 is related to ring cell inflation, vegetative growth, and conidiation. • DdaSTE12 deletion resulted in defects in trap formation and ring development.

THRIVE, ASTRAL, and iScore scales for predicting prognosis of mechanical thrombectomy in patients with acute ischemic stroke.

OBJECTIVE: To validate the predictive performance of the THRIVE, ASTRAL, and iScore scales for clinical functional outcomes following mechanical thrombectomy (MT) for acute ischemic stroke (AIS). METHODS: A total of 111 patients meeting the inclusion criteria were included in this study, with 59 (53.2%) having a good prognosis and 52 (46.8%) having a poor prognosis. MedCalc software was applied to plot receiver operating characteristic (ROC) curves, calculate the area under the curve (AUC), and compare the predictive efficacy of the three scales two by two using Delong text. Statistical significance was defined as Pc < 0.05. RESULTS: Logistic binary regression multifactorial analysis revealed that iScore is one of the poor predictors of prognosis in patients with MT. The AUC values for the THRIVE, ASTRAL, and iScore scales in predicting prognosis after MT were found to be 0.713, 0.738, and 0.820, respectively. CONCLUSION: The iScore is a reliable tool for assessing the poor prognosis of MT in patients with AIS.

Clinical study of the prognostic value of the Essen score for acute cerebral infarction.

OBJECTIVE: A few clinical studies have been conducted on the prognostic value of the Essen score in acute cerebral infarction (ACI), and this study explores whether the Essen score can assess the prognosis of ACI. METHODS: Data were collected from 1176 patients with ACI. The patients were divided into three groups on the basis of the Essen score, with groups 1, 2 and 3 having scores of 0-2, 3-6 and 7-9, respectively. Logistic multivariate analysis was performed to analyse the predictors of poor prognosis in patients with ACI. The X2 trend test was used to compare the poor-prognosis groups on the basis of the Essen score. The receiver operating characteristic (ROC) curve of patient prognosis was plotted using MedCalc software, and the area under the ROC curve (AUC) was calculated. P < 0.05 was considered statistically significant. RESULTS: Multivariate analysis of the good- and poor-prognosis groups of ACI showed that the Essen score and the male gender were predictors of poor prognosis. The X2 trend test was used to compare the poor-prognosis groups on the basis of the Essen score, and results suggested that the higher the Essen score was, the worse the prognosis was. The Essen score assessed the prognosis of ACI with an AUC of 0.787 and P < 0.001. CONCLUSION: The Essen score is a valuable scoring system for predicting the prognosis of patients with ACI.

Relationship between maximum carotid plaque area and prognosis of patients with acute ischaemic stroke.

OBJECTIVE: In this study, the relationship between maximum carotid plaque area and stroke prognosis was analysed by carotid ultrasonography, and the relevant risk factors affecting the prognosis of acute ischaemic stroke (AIS) were investigated to provide novel insights into stroke prevention and management. METHODS: A total of 205 AIS patients with carotid plaques were included in this study. Based on the mRS score at discharge, patients with AIS were classified into the good prognosis group (mRS ≤ 2) and poor prognosis group (mRS ≥ 3). SPSS 25.0 was used to analyse the data. Univariate and multivariate analyses were performed on the two groups with good and poor prognosis. Comparison between good and poor prognosis in patients with AIS in different circulatory systems was performed using the Kruskal-Wallis test. Differences were considered statistically significant at P < 0.05. RESULTS: Comparison of baseline data revealed differences in carotid plaque diameter, carotid intima-media thickness, maximum carotid plaque area, history of previous stroke and plaque echogenicity between the good and poor prognosis groups (P < 0.05). Results of multifactorial analyses of logistic binary regression indicated that history of previous stroke and maximum carotid plaque area were predictors of poor prognosis, with odds ratio of 2.515 (95%CI [1.327-4.764]) and 1.019 (95%CI [1.006-1.032]), respectively. CONCLUSION: The maximum carotid plaque area and history of previous stroke are important predictors for assessing prognosis in patients with AIS.

Barthel Index, SPAN-100, and NIHSS Studies on the Predictive Value of Prognosis in Patients With Thrombolysis.

OBJECTIVE: We mainly explore the predictive value of Barthel Index (BI), SPAN-100, and National Institute of Health stroke scale (NIHSS) scores on clinical prognosis and functional outcomes in thrombolytic patients and compare the differences in the predictive values of the above 3 scales so as to provide an effective basis to evaluate the prognosis of thrombolytic patients. METHODS: Data were collected from 212 patients with the first-onset AIS (acute ischemic stroke). The enrolled patients were treated with recombinant tissue plasminogen activator thrombolytic therapy and were divided into 2 groups according to the modified Rankin Scale (mRS) score at discharge: the prognosis group (mRS≤2 points) and the poor prognosis group (mRS≥3 points). Logistic multivariate analysis was used to analyze the predictors of poor prognosis in patients with thrombolysis. MedCalc software was used to plot receiver operating characteristic (ROC) curves, calculate the area under the ROC curve (AUC), and compare the prediction performance of the 3 scales by the Delong and colleagues' method, and the difference of P <0.05 was statistically significant. RESULTS: Logistic binary regression multivariate analysis suggested that BI was a predictor of poor prognosis for thrombolytic therapy in patients with AIS. The lower the BI score, the poorer the prognosis. The AUC for BI score was 0.862, 95% CI (0.808-0.906), NIHSS score AUC was 0.665, 95% CI (0.597-0.728), and SPAN-100 score AUC was 0.640, 95% CI (0.572-0.705). AUC comparison of 3 scoring ROC curves suggested statistically significant differences between BI and NIHSS ( PC =0.0000), BI and SPAN-100 ( PC =0.0000); no significant difference was observed between SPAN-100 and NIHSS ( PC =1.7997). CONCLUSIONS: Simple BI scores have a high prognostic value for thrombolytic therapy in AIS.

Characterization of genome-wide phylogenetic conflict uncovers evolutionary modes of carnivorous fungi.

Mass extinction has often paved the way for rapid evolutionary radiation, resulting in the emergence of diverse taxa within specific lineages. The emergence and diversification of carnivorous nematode-trapping fungi (NTF) in Ascomycota have been linked to the Permian-Triassic (PT) extinction, but the processes underlying NTF radiation remain unclear. We conducted phylogenomic analyses using 23 genomes that represent three NTF lineages, each employing distinct nematode traps-mechanical traps (Drechslerella spp.), three-dimensional (3D) adhesive traps (Arthrobotrys spp.), and two-dimensional (2D) adhesive traps (Dactylellina spp.), and the genome of one non-NTF species as the outgroup. These analyses revealed multiple mechanisms that likely contributed to the tempo of the NTF evolution and rapid radiation. The species tree of NTFs based on 2,944 single-copy orthologous genes suggested that Drechslerella emerged earlier than Arthrobotrys and Dactylellina. Extensive genome-wide phylogenetic discordance was observed, mainly due to incomplete lineage sorting (ILS) between lineages. Two modes of non-vertical evolution (introgression and horizontal gene transfer) also contributed to phylogenetic discordance. The ILS genes that are associated with hyphal growth and trap morphogenesis (e.g., those associated with the cell membrane system and polarized cell division) exhibited signs of positive selection.IMPORTANCEBy conducting a comprehensive phylogenomic analysis of 23 genomes across three NTF lineages, the research reveals how diverse evolutionary mechanisms, including ILS and non-vertical evolution (introgression and horizontal gene transfer), contribute to the swift diversification of NTFs. These findings highlight the complex evolutionary dynamics that drive the rapid radiation of NTFs, providing valuable insights into the processes underlying their diversity and adaptation.

The mycobiome: interactions with host and implications in diseases.

Over the past decade, our understanding of the composition and function of the human mucosal surface-associated fungal community (i.e. the mycobiome) has rapidly expanded. Fungi colonize at various sites of the mucosal surface at birth and play important roles in the development and homeostasis of immune system throughout adulthood. Here, we review the recent research progresses in the human mycobiome at different body sites, including the gastrointestinal (GI) tract, the respiratory tract, the urogenital tract, the oral cavity, the skin surface, and the tumor tissues. Researchers have made extensive effort in characterizing the interactions between mycobiome and immune system, especially in the GI tract. We discuss the mycobiome dysbiosis and its implications to the progression of diseases such as inflammatory bowel diseases, alcoholic liver diseases, systemic infections, cancers, and so on, indicating the potential of mycobiome-targeting intervention strategy for life-threatening diseases.

Multiple Roles of the Low-Affinity Calcium Uptake System in Drechslerella dactyloides, a Nematode-Trapping Fungus That Forms Constricting Rings.

(1) Background: the low-affinity calcium uptake system (LACS) has been shown to play a crucial role in the conidiation and formation of adhesive nets and knobs by nematode-trapping fungi (NTF), but its involvement in the formation of constricting rings (CRs), mechanical traps to capture free-living nematodes, remains unexplored. (2) Methods: we investigated the function of two LACS genes (DdaFIG_1 and DdaFIG_2) in Drechslerella dactyloides, an NTF that forms CRs. We generated single (DdaFIG_1Ri and DdaFIG_2Ri) and double (DdaFIG_1,2Ri) knockdown mutants via the use of RNA interference (RNAi). (3) Results: suppression of these genes significantly affected conidiation, trap formation, vegetative growth, and response to diverse abiotic stresses. The number of CRs formed by DdaFIG_1Ri, DdaFIG_2Ri, and DdaFIG_1,2Ri decreased to 58.5%, 59.1%, and 38.9% of the wild-type (WT) level, respectively. The ring cell inflation rate also decreased to 73.6%, 60.6%, and 48.8% of the WT level, respectively. (4) Conclusions: the LACS plays multiple critical roles in diverse NTF.

A palisade-shaped membrane reservoir is required for rapid ring cell inflation in Drechslerella dactyloides.

Fusion of individual vesicles carrying membrane-building materials with the plasma membrane (PM) enables gradual cell expansion and shape change. Constricting ring (CR) cells of carnivorous fungi triple in size within 0.1-1 s to capture passing nematodes. Here, we investigated how a carnivorous fungus, Drechslerella dactyloides, executes rapid and irreversible PM expansion during CR inflation. During CR maturation, vesicles carrying membrane-building materials accumulate and fuse, forming a structure named the Palisade-shaped Membrane-building Structure (PMS) around the rumen side of ring cells. After CR inflation, the PMS disappears, with partially inflated cells displaying wavy PM and fully inflated cells exhibiting smooth PM, suggesting that the PMS serves as the reservoir for membrane-building materials to enable rapid and extensive PM expansion. The DdSnc1, a v-SNARE protein, accumulates at the inner side of ring cells and is necessary for PMS formation and CR inflation. This study elucidates the unique cellular mechanisms underpinning rapid CR inflation.

The mechanism of formononetin/calycosin compound optimizing the effects of temozolomide on C6 malignant glioma based on metabolomics and network pharmacology.

The complex of formononetin and calycosin (FMN/CAL) shows a synergistic effect on temozolomide in the treatment of malignant glioma, however the mechanism is unclear. We investigated the mechanism through means of metabolomics, network pharmacology and molecular biology. FMN/CAL enhanced the inhibition of TMZ on the growth and infiltration of C6 glioma. The metabolomic results showed that the TMZ sensitization of FMN/CAL mainly involved 5 metabolic pathways and 4 metabolites in cells, 1 metabolic pathway and 2 metabolites in tumor tissues, and 7 metabolic pathways and 8 metabolites in serum. Further network pharmacological analysis revealed that NOS2 was a potential target for FMN/CAL to regulate the metabolism in TMZ-treated C6 glioma cells, serums and tissues, and TNF-α was another potential target identified in tissues. FMN/CAL down-regulated the expression of NOS2 in tumor cells and tissues, and reduced the secretion of TNF-α in tumor region. FMN/CAL promoted TMZ-induced C6 cell apoptosis by inhibiting NOS2, but the inhibition of cell vitality and migration was not through NOS2. Our work revealed that FMN/CAL can increase the sensitivity of malignant glioma to TMZ by inhibiting NOS2-dependent cell survival, which provides a basis for the application of this combination in adjuvant treatment of glioma.

DdaCrz1, a C2H2-Type Transcription Factor, Regulates Growth, Conidiation, and Stress Resistance in the Nematode-Trapping Fungus Drechslerella dactyloides.

The Ca2+/calmodulin-dependent signaling pathway regulates diverse cellular processes. Calcineurin is a calcium-dependent phosphatase acting in fungi mainly through Crz1, a zinc finger transcription factor. Although the likely involvement of Ca2+ in fungal carnivorism has been documented, how Crz1 functions in nematode-trapping fungi remains unknown. Here, we identified the Crz1 gene (named as DdaCrz1) in Drechslerella dactyloides, a species that forms constricting rings to trap nematodes. The deletion of DdaCrz1 significantly reduced hyphal growth and conidiation, trap formation, and ring cell inflation. Moreover, the mutation increased sensitivity to Mn2+ but decreased sensitivity to Ca2+, Mg2+, Zn2+, and Li+. Similarly, the mutant showed increased tolerance to osmotic stress but was more sensitive to Congo red, a cell wall-damaging agent. Our results confirmed the critical roles of the Ca2+/calmodulin-dependent signaling pathway in regulating growth, conidiation, and the stress response, and suggested its involvement in trapping nematodes.

SNARE Protein DdVam7 of the Nematode-Trapping Fungus Drechslerella dactyloides Regulates Vegetative Growth, Conidiation, and the Predatory Process via Vacuole Assembly.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins play conserved roles in membrane fusion events in eukaryotes and have been documented to be involved in fungal growth and pathogenesis. However, little is known about the roles of SNAREs in trap morphogenesis in nematode-trapping fungi (NTF). Drechslerella dactyloides, one of the constricting ring-forming NTF, captures free-living nematodes via rapid ring cell inflation. Here, we characterized DdVam7 of D. dactyloides, a homolog of the yeast SNARE protein Vam7p. Deletion of DdVam7 significantly suppressed vegetative growth and conidiation. The mutation significantly impaired trap formation and ring cell inflation, resulting in a markedly decreased nematode-trapping ability. A large vacuole could develop in ring cells within ~2.5 s after instant inflation in D. dactyloides. In the ΔDdVam7 mutant, the vacuoles were small and fragmented in hyphae and uninflated ring cells, and the large vacuole failed to form in inflated ring cells. The localization of DdVam7 in vacuoles suggests its involvement in vacuole fusion. In summary, our results suggest that DdVam7 regulates vegetative growth, conidiation, and the predatory process by mediating vacuole assembly in D. dactyloides, and this provides a basis for studying mechanisms of SNAREs in NTF and ring cell rapid inflation. IMPORTANCE D. dactyloides is a nematode-trapping fungus that can capture nematodes through a constricting ring, the most sophisticated trapping device. It is amazing that constricting ring cells can inflate to triple their size within seconds to capture a nematode. A large centrally located vacuole is a unique signature associated with inflated ring cells. However, the mechanism underpinning trap morphogenesis, especially vacuole dynamics during ring cell inflation, remains unclear. Here, we documented the dynamics of vacuole assembly during ring cell inflation via time-lapse imaging for the first time. We characterized a SNARE protein in D. dactyloides (DdVam7) that was involved in vacuole assembly in hyphae and ring cells and played important roles in vegetative growth, conidiation, trap morphogenesis, and ring cell inflation. Overall, this study expands our understanding of biological functions of the SNARE proteins and vacuole assembly in NTF trap morphogenesis and provides a foundation for further study of ring cell rapid inflation mechanisms.

The E3-ligase AoUBR1 in N-end rule pathway is involved in the vegetative growth, secretome, and trap formation in Arthrobotrys oligospora.

The N-end rule pathway is a regulated protein degradation system. Arthrobotrys oligospora, a typical nematode-trapping fungus, switches its life strategies from saprophytism to carnivorism when capturing free-living nematodes by means of adhesive networks. In this study, a putative E3-ligase AoUBR1 involved in N-end rule pathway was characterized in A. oligospora during vegetative growth and trap formation. Expression of AoUBR1 coding gene was down-regulated during trap formation. Compared with wild type, the AoUBR1 knock-out mutants decreased the vegetative growth, formed less traps, and turned to be sensitive to cold stress, while, AoUBR1 overexpression mutants lost the capacity to produce conidia and also formed less traps. A number of genes differentially expressed by knock-out and overexpression of AoUBR1, which lead to the transcriptional responses associated with plasma membrane, transportation, oxidation, and proteolysis. AoUBR1 knock-out also resulted in the down-regulation of numerous secreted proteins associated with carnivorism and nutrient utilization from nematodes. In addition, AoUBR1 homologs were conserved in nematode-trapping fungi based on the genome searching. Therefore, the results suggested AoUBR1 in A. oligospora and its homologs in other trapping fungi are involved in the lifestyle switch between saprophytism and carnivorism.

Proteomic analysis of healthy and atretic porcine follicular granulosa cells.

Follicular atresia is initiated with the apoptosis of granulosa cells (GCs) after birth in mammals. The molecular mechanisms underlying GC apoptosis during follicular selection are unclear at present. The objective of this study is to identify the proteins and pathways that may be involved in porcine follicular atresia. Proteins isolated from GCs collected from healthy and atretic follicles were detected by tandem mass tag (TMT) protein labeling and LC-MS/MS. A total of 4591 proteins in the healthy follicle granulosa cell (HFGC) and atretic follicle granulosa cell (AFGC) groups were identified, and 399 differentially abundant proteins were found between the HFGC and AFGC groups; of which 262 proteins were significantly up-regulated and 137 proteins were significantly down-regulated. Differential protein enrichment analysis showed that proteins involved in proteolysis, protein destabilization, phagocytosis, and engulfment were more abundant in the AFGC group. Also, these proteins were mainly involved in the lysosome, phagosome, autophagy, and apoptosis pathways. Specially, PTGFRN is potential important regulated protein in the development of the antral follicle, and down-regulation of PTGFRN in GCs may lead to follicular atresia. Our study shows that the identified proteins and their related signaling pathways may play crucial roles during health follicle develop to atretic follicle. SIGNIFICANCE: Follicular atresia during 'selection' reduces the reproductive potential of sows. In this study, we found 399 proteins differentially abundant. between the HFGC and AFGC groups. These results establish a foundation for elucidating the mechanism of follicular atresia in swine.

Evaluation of Strategies for Decreasing Blood Glucose Using Albuminbinding Domain.

OBJECTIVE: Frequent administrations for DPPIV-resistant GLP-1 analogs are necessary to maintain the blood concentrations due to the short half-life of less than 5 minutes. However, most delivery systems that possess the ability of sustainable release of GLP-1 have drawbacks such as low yield, high cost and undesirable side effects. Therefore, we aimed to prepare a simple and efficient delivery system that could be feasibly applied to reduce blood glucose. METHODS: A novel GLP-1 delivery system (GLP-1-ELPs-SA) was prepared and characterized by circular dichroism. Furthermore, the activity and property of GLP-1-ELPs-SA were evaluated in vitro and in vivo. RESULTS: GLP-1-ELPs-SA are easily expressed in E. coli in a soluble formulation and purified through the inverse transition cycle. GLP-1-ELPs-SA spontaneously generated depot under physiological conditions. GLP-1-ELPs-SA was also found to be dispersed in the blood vessels from the depot and showed a high affinity to bind with mice (C57BL/6J) albumin, which shows that GLP-1-ELPs-SA has a long circulation time in vivo. CONCLUSION: Our delivery system could markedly decrease the clearance of recombinant proteins based on serum albumin, without substantially increasing the protein molecular weight and remarkably reducing the blood glucose within 120 h.

In vitro and in vivo Study on Glioma Treatment Enhancement by Combining Temozolomide with Calycosin and Formononetin.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragalina alpestris is a traditional Chinese herbal medicine with anti-inflammatory, anti-immune, anti-tumor and other pharmacological effects. Calycosin and formononetin (FMN) are two natural compounds isolated from astragalus. It has been shown that calycosin and FMN are active anti-tumor ingredient. AIM OF THE STUDY: The aim of this current work was to study the therapeutic enhancement of temozolomide (TMZ) on gliomavia combining with calycosin and FMN. MATERIALS AND METHODS: The effect of co-administration via hematoxylin-eosin staining (HE staining) was determined by measuring cell proliferation toxicity with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, Thiazolyl Blue Tetrazolium Bromide (MTT) assay and sequentially observing the cell morphology. To synchronously explore the effect of migration on C6, transwell assay and wound healing assay were performed. Apoptosis was measured by Annexin V/propidium iodide (PI) staining. Meanwhile, western blot was used to investigate proteins involved in the mechanisms for migration and apoptosis. Furthermore, HE staining and immunohistochemistry were also analyzed for curative effect in vivo. RESULTS: The efficacy of TMZ on glioma could be enhanced by combining with calycosin and FMN through inhibiting the proliferation and migration of glioma cells and promoting their apoptosis. Western blot assays indicated that expression of apoptotic proteins (Bcl-2 Associated XProtein (Bax), Cleaved Caspase-3, Cleaved Caspase-9) were up-regulated. Anti-apoptotic protein (B-cell lymphoma-2,Bcl-2) was down-regulated. The migratory proteins (Matrix metallopeptidase 2, 9, MMP-2, MMP-9) was downregulated. In vivo study, this kind of co-administration (calycosin, FMN, and TMZ) exhibited the marked therapeutic effect on glioma. CONCLUSIONS: This study has identified that calycosin and FMN can increase the treatment effect of TMZ in vitro and in vivo. These attractive features substantially broadened the application range of TMZ as a glioma treatment medicine.