Dual-Atom Nanozyme Eye Drops Attenuate Inflammation and Break the Vicious Cycle in Dry Eye Disease.

Dry eye disease (DED) is a major ocular pathology worldwide, causing serious ocular discomfort and even visual impairment. The incidence of DED is gradually increasing with the high-frequency use of electronic products. Although inflammation is core cause of the DED vicious cycle, reactive oxygen species (ROS) play a pivotal role in the vicious cycle by regulating inflammation from upstream. Therefore, current therapies merely targeting inflammation show the failure of DED treatment. Here, a novel dual-atom nanozymes (DAN)-based eye drops are developed. The antioxidative DAN is successfully prepared by embedding Fe and Mn bimetallic single-atoms in N-doped carbon material and modifying it with a hydrophilic polymer. The in vitro and in vivo results demonstrate the DAN is endowed with superior biological activity in scavenging excessive ROS, inhibiting NLRP3 inflammasome activation, decreasing proinflammatory cytokines expression, and suppressing cell apoptosis. Consequently, the DAN effectively alleviate ocular inflammation, promote corneal epithelial repair, recover goblet cell density and tear secretion, thus breaking the DED vicious cycle. Our findings open an avenue to make the DAN as an intervention form to DED and ROS-mediated inflammatory diseases.

Intelligent Vascularized 3D/4D/5D/6D-Printed Tissue Scaffolds.

Blood vessels are essential for nutrient and oxygen delivery and waste removal. Scaffold-repairing materials with functional vascular networks are widely used in bone tissue engineering. Additive manufacturing is a manufacturing technology that creates three-dimensional solids by stacking substances layer by layer, mainly including but not limited to 3D printing, but also 4D printing, 5D printing and 6D printing. It can be effectively combined with vascularization to meet the needs of vascularized tissue scaffolds by precisely tuning the mechanical structure and biological properties of smart vascular scaffolds. Herein, the development of neovascularization to vascularization to bone tissue engineering is systematically discussed in terms of the importance of vascularization to the tissue. Additionally, the research progress and future prospects of vascularized 3D printed scaffold materials are highlighted and presented in four categories: functional vascularized 3D printed scaffolds, cell-based vascularized 3D printed scaffolds, vascularized 3D printed scaffolds loaded with specific carriers and bionic vascularized 3D printed scaffolds. Finally, a brief review of vascularized additive manufacturing-tissue scaffolds in related tissues such as the vascular tissue engineering, cardiovascular system, skeletal muscle, soft tissue and a discussion of the challenges and development efforts leading to significant advances in intelligent vascularized tissue regeneration is presented.

Deep-Learning Terahertz Single-Cell Metabolic Viability Study.

Cell viability assessment is critical, yet existing assessments are not accurate enough. We report a cell viability evaluation method based on the metabolic ability of a single cell. Without culture medium, we measured the absorption of cells to terahertz laser beams, which could target a single cell. The cell viability was assessed with a convolution neural classification network based on cell morphology. We established a cell viability assessment model based on the THz-AS (terahertz-absorption spectrum) results as y = a = (x - b)c, where x is the terahertz absorbance and y is the cell viability, and a, b, and c are the fitting parameters of the model. Under water stress the changes in terahertz absorbance of cells corresponded one-to-one with the apoptosis process, and we propose a cell 0 viability definition as terahertz absorbance remains unchanged based on the cell metabolic mechanism. Compared with typical methods, our method is accurate, label-free, contact-free, and almost interference-free and could help visualize the cell apoptosis process for broad applications including drug screening.

Plant exosome nanovesicles (PENs): green delivery platforms.

Natural plants have been attracting increasing attention in biomedical research due to their numerous benefits. Plant exosome-derived vesicles, some of the plant's components, are small nanoscale vesicles secreted by plant cells. These vesicles are rich in bioactive substances and play significant roles in intercellular communication, information transfer, and maintaining homeostasis in organisms. They also hold promise for treating diseases, and their vesicular structures make them suitable carriers for drug delivery, with large-scale production feasible. Therefore, this paper aims to provide an overview of nanovesicles from different plant sources and their extraction methods. We also outline the biological activities of nanovesicles, including their anti-inflammatory, anti-viral, and anti-tumor properties, and systematically introduce their applications in drug delivery. These applications include transdermal delivery, targeted drug delivery, gene delivery, and their potential use in the modern food industry. This review provides new ideas and methods for future research on plant exosomes, including their empowerment by artificial intelligence and gene editing, as well as their potential application in the biomedicine, food, and agriculture industries.

Hazardous Gases-Responsive Photonic Crystals Cryogenic Sensors Based on Antifreezing and Water Retention Hydrogels.

Nowadays, the sensing of hazardous gases is urgent for the consideration of public safety and human health, especially in extreme conditions of low temperatures. In this study, a photonic crystals (PhCs) sensor with water retention and antifreezing properties was developed and applied to visual hazardous gases sensing at low temperature, passively. The sensor was prepared by dip-coating with poly(methyl methacrylate) (PMMA) colloidal microspheres followed by embedding in k-carrageenan/polyacrylamide-ethylene glycol (k-CA/PAM-EG) hydrogel. The sensor responded to hazardous gases, including ammonia, toluene, xylene, acetone, methanol, ethanol, and 1-propanol, with a change in the reflection wavelength and visible structural color. At room temperature, the reflection wavelength of the sensor blue-shifted 49 nm in ammonia, and the structural color changed from red to yellow. For low temperatures, the sensor showed great water retention and antifreezing properties even at -57 °C due to the double network. The sensor still had a great response to hazardous gases after freezing at -20 °C for 12 h and testing at 0 °C, and the obtained results were similar to those at room temperature. Based on this excellent stability and visual sensing at low temperature, the sensor demonstrates the potential for detection of hazardous vapors in extreme environments.

Doublecortin-Expressing Neurons in Human Cerebral Cortex Layer II and Amygdala from Infancy to 100 Years Old.

A cohort of morphologically heterogenous doublecortin immunoreactive (DCX +) "immature neurons" has been identified in the cerebral cortex largely around layer II and the amygdala largely in the paralaminar nucleus (PLN) among various mammals. To gain a wide spatiotemporal view on these neurons in humans, we examined layer II and amygdalar DCX + neurons in the brains of infants to 100-year-old individuals. Layer II DCX + neurons occurred throughout the cerebrum in the infants/toddlers, mainly in the temporal lobe in the adolescents and adults, and only in the temporal cortex surrounding the amygdala in the elderly. Amygdalar DCX + neurons occurred in all age groups, localized primarily to the PLN, and reduced in number with age. The small-sized DCX + neurons were unipolar or bipolar, and formed migratory chains extending tangentially, obliquely, and inwardly in layers I-III in the cortex, and from the PLN to other nuclei in the amygdala. Morphologically mature-looking neurons had a relatively larger soma and weaker DCX reactivity. In contrast to the above, DCX + neurons in the hippocampal dentate gyrus were only detected in the infant cases in parallelly processed cerebral sections. The present study reveals a broader regional distribution of the cortical layer II DCX + neurons than previously documented in human cerebrum, especially during childhood and adolescence, while both layer II and amygdalar DCX + neurons persist in the temporal lobe lifelong. Layer II and amygdalar DCX + neurons may serve as an essential immature neuronal system to support functional network plasticity in human cerebrum in an age/region-dependent manner.

Specific Alcohol-Responsive Photonic Crystal Sensors Based on Host-Guest Recognition.

A photonic crystal material based on ß-cyclodextrin (ß-CD) with adsorption capacity is reported. The materials ((A-ß-CD)-AM PC) consist of 3D poly (methyl methacrylate) (PMMA) colloidal microsphere arrays and hydrogels supplemented with ß-cyclodextrin modified by acryloyl chloride. The prepared materials are then utilized for VOCs gas sensing. The 3D O-(A-ß-CD)-AM PC was used to detect toluene, xylene, and acetone and the response was seen as the red-shift of the reflection peak. The 3D I-(A-ß-CD)-AM PC was used to detect toluene, xylene, and acetone which occurred redshifted, while methanol, ethanol, and propanol and the peaks' red-shifting was observed. However, among these, methanol gave the largest red-shift response The sensor has broad prospects in the detection of alcohol and the detection of alcohol-loaded drug releases in the future.

Health Risk Assessment of Inhalation Exposure to Airborne Particle-Bound Nitrated Polycyclic Aromatic Hydrocarbons in Urban and Suburban Areas of South China.

Airborne particulates (PM2.5 and TSP) were collected from outdoor and indoor areas at urban (Haizhu District) and suburban (Huadu District) sites from 2019 to 2020 in Guangzhou. Three nitro-polycyclic aromatic hydrocarbons (nitro-PAHs) in the airborne particulates were identified by a gas chromatograph equipped with a triple-quadrupole mass spectrometer. In the Haizhu District and Huadu District, the nitro-PAH concentrations in PM2.5 and TSP did not show a significant decrease from winter to summer. From 2019 to 2020, the difference in the average concentration of nitro-PAHs in PM2.5 and TSP in Guangzhou was relatively low and had no statistical significance. The diagnostic ratios of 2-nitrofluorene (2-NF)/1-nitropyrene (1-NP) in TSP are less than five, while for 2-NF/1-NP in outdoor PM2.5 in the summer of 2019 and 2020 are more than five, which indicates that nitro-PAHs in the atmospheric PM2.5 in Guangzhou during summer mainly originated from the secondary formation of atmospheric photochemical reactions between parent PAHs and oxidants (·OH, NO3, and O3). 9-Nitroanthracene (9-NT) made the most significant contribution to the total nitro-PAH concentration. The incremental lifetime cancer risks (ILCRs) of nitro-PAHs in PM2.5 and TSP by inhalation exposure indicated low potential health risks in the urban-suburban of Guangzhou.

Intraneuronal sortilin aggregation relative to granulovacuolar degeneration, tau pathogenesis and sorfra plaque formation in human hippocampal formation.

Extracellular ß-amyloid (Aß) deposition and intraneuronal phosphorylated-tau (pTau) accumulation are the hallmark lesions of Alzheimer's disease (AD). Recently, "sorfra" plaques, named for the extracellular deposition of sortilin c-terminal fragments, are reported as a new AD-related proteopathy, which develop in the human cerebrum resembling the spatiotemporal trajectory of tauopathy. Here, we identified intraneuronal sortilin aggregation as a change related to the development of granulovacuolar degeneration (GVD), tauopathy, and sorfra plaques in the human hippocampal formation. Intraneuronal sortilin aggregation occurred as cytoplasmic inclusions among the pyramidal neurons, co-labeled by antibodies to the extracellular domain and intracellular C-terminal of sortilin. They existed infrequently in the brains of adults, while their density as quantified in the subiculum/CA1 areas increased in the brains from elderly lacking Aß/pTau, with pTau (i.e., primary age-related tauopathy, PART cases), and with Aß/pTau (probably/definitive AD, pAD/AD cases) pathologies. In PART and pAD/AD cases, the intraneuronal sortilin aggregates colocalized partially with various GVD markers including casein kinase 1 delta (Ck1δ) and charged multivesicular body protein 2B (CHMP2B). Single-cell densitometry established an inverse correlation between sortilin immunoreactivity and that of Ck1δ, CHMP2B, p62, and pTau among pyramidal neurons. In pAD/AD cases, the sortilin aggregates were reduced in density as moving from the subiculum to CA subregions, wherein sorfra plaques became fewer and absent. Taken together, we consider intraneuronal sortilin aggregation an aging/stress-related change implicating protein sorting deficit, which can activate protein clearance responses including via enhanced phosphorylation and hydrolysis, thereby promoting GVD, sorfra, and Tau pathogenesis, and ultimately, neuronal destruction and death.

Chiral gold clusters functionalized two-dimensional nanoparticle films to regulate the adhesion and differentiation of stem cells.

Chirality has been proved to play a significant role in tuning cell behaviors and controlling cellular functions. Up to now, almost all the chirality origins of extracellular microenvironment are belong to chiral ligands induction or direct chiral patterns. In this study, chiral gold nanoclusters (L/D-AuNC) loaded on two-dimensional gold nanoparticle films (L/D-film) with multiple chirality origins were prepared to regulate the adhesion and differentiation of mouse bone marrow mesenchymal stem cells (MSCs). MSCs on the D-film exhibited higher cell density and larger spreading area, and more cells differentiated into osteoblasts. Compared with D-film, L-film has a lower cell density and smaller spreading area, and more adipoblasts are achieved. The corresponding expression results of osteogenic differentiation marker (RUNX2) also confirmed the above experimental phenomenon. These results demonstrated that the chirality of clusters has great effect on the direction of cell fate.

Dual-Responsive Photonic Crystal Sensors Based on Physical Crossing-Linking SF-PNIPAM Dual-Crosslinked Hydrogel.

Flexible wearable materials have frequently been used in drug delivery, healthcare monitoring, and wearable sensors for decades. As a novel type of artificially designed functional material, photonic crystals (PCs) are sensitive to the changes in the external environment and stimuli signals. However, the rigidity of the PCs limits their application in the field of biometric and optical sensors. This study selects silk fibroin (SF) and poly-N-isopropylacrylamide (PNIPAM) as principal components to prepare the hydrogel with the physical crosslinking agent lithium silicate (LMSH) and is then integrated with PCs to obtain the SF-PNIPAM dual-crosslinked nanocomposite for temperature and strain sensing. The structural colors of the PCs change from blue to orange-red by the variation in temperature or strain. The visual temperature-sensing and adhesion properties enable the SF-PNIPAM dual-crosslinked nanocomposite to be directly attached to the skin in order to monitor the real-time dynamic of human temperature. Based on its excellent optical properties and biocompatibility, the SF-PNIPAM dual-crosslinked nanocomposite can be applied to the field of visual biosensing, wearable display devices, and wound dressing materials.

Transcriptome-Wide m6A Methylome and m6A-Modified Gene Analysis in Asthma.

N6-methyladenosine (m6A) modification is one of the most prevalent RNA modification forms and is an important posttranscriptional mechanism for regulating genes. In previous research, we found that m6A regulator-mediated RNA methylation modification was involved in asthma; however, the specific modified genes are not clear. In this study, we systematically evaluated the transcriptome-wide m6A methylome and m6A-modified genes in asthma. Here, we performed two high-throughput sequencing methods, methylated RNA immunoprecipitation sequencing (MeRIP-seq), and RNA sequencing (RNA-seq) to identify key genes with m6A modification in asthma. Through difference analysis, we found that 416 methylation peaks were significantly upregulated and 152 methylation peaks were significantly downregulated, and it was mainly distributed in 3' UTR. Furthermore, compared with the control group, there were 2,505 significantly upregulated genes and 4,715 significantly downregulated genes in the asthma group. Next, through a combined analysis of transcriptome and differential peaks, 14 differentially expressed genes related to RNA methylation modification were screened. Finally, through 87 health controls and 411 asthma cases from the U-BIOPRED (Unbiased Biomarkers for the Prediction of Respiratory Disease Outcomes) program, we verified three m6A-modified key genes (BCL11A, MATK, and CD300A) and found that they were mainly distributed in exons and enriched in 3' UTR. Our findings suggested that intervening in m6A-modified genes may provide a new idea for the treatment of asthma.

Long Non-coding RNA Double Homeobox A Pseudogene 8: A Novel Oncogenic Propellant in Human Cancer.

A growing number of studies are reporting important roles played by long non-coding RNAs (lncRNAs) in various pathological and physiological processes. LncRNAs are implicated in numerous genomic regulatory functions at different levels, including regulation of transcription, post-transcriptional processes, genomic stability, and epigenetic genome modifications. Double homeobox A pseudogene 8 (DUXAP8), a novel lncRNA, has been reported to be involved in many cancers, including gastric, colorectal, esophageal, bladder, oral, ovarian, lung, and pancreatic cancers as well as hepatocellular carcinoma (HCC). DUXAP8 plays specific oncogenic roles via numerous malignancies promoting pathways. DUXAP8 is frequently dysregulated in multiple cancers, acting as a sponge to downregulate various tumor-suppressing microRNA activities. In this review, we comprehensively explore DUXAP8 expression and prognosis across cancer types, and systematically summarize current evidence concerning the functions and molecular mechanisms of DUXAP8 in tumorigenesis and progression. We conclude that DUXAP8 is a potential biomarker and therapeutic target for multiple cancers.

Advances in poly(rC)-binding protein 2: Structure, molecular function, and roles in cancer.

Poly(rC)-binding protein 2 (PCBP2) is an RNA-binding protein that is characterized by its ability to interact with poly(C) with high affinity in a sequence-specific manner. PCBP2 contains three K homology domains, which are consensus RNA-binding domains that play a role in recognizing and combining with RNA and DNA. The specific structure and localization of PCBP2 lay the foundation for its multiple roles in transcriptional, posttranscriptional, and translational processes, even in iron metabolism. Numerous studies have indicated that PCBP2 expression is increased in many cancer types. PCBP2 is considered as an oncogene that promotes tumorigenesis, development of cancer cells, and metastasis. Here, we summarized the current evidence regarding PCBP2 in the proliferation, migration, invasion of cancer cells, and drug resistance, aiming to clarify the molecular mechanisms of PCBP2 in cancer. Results from this review suggest that an in-depth study of PCBP2 in cancer may provide novel biomarkers for prognostic or therapeutic purposes.

Aqueous extract of Taxus chinensis var. mairei regulates the Hippo-YAP pathway and promotes apoptosis of non-small cell lung cancer via ATF3 in vivo and in vitro.

Taxus chinensis var. mairei (TC) is a traditional Chinese ornamental and medicinal plant, the leaves and twigs of which are used in anti-tumor therapy in southern China. However, the mechanism and role of aqueous extract of TC (AETC) in promoting apoptosis in non-small cell lung cancer (NSCLC) cell lines has remained unclear. In this research, we observed that AETC inhibited the suppression of the proliferation of NSCLC cells and highly inhibited the proliferation of NCI-1975 cells. Furthermore, AETC exerted minimal inhibitory effects on normal human lung epithelial cells and induced apoptosis in NCI-1975 and A549 cells. The findings of RNA sequencing, qRT-PCR, western blotting, and immunofluorescence showed that upregulated ATF3 expression and ATF3 gene knockdown, respectively, increased and decreased the anti-tumor effects of AETC associated with Hippo pathway inhibition and decreased YAP degradation. Furthermore, AETC reduced the tumor volume and weight in nude mice; upregulated ATF3, p-MOB1, and p-YAP (Ser397); and actively regulated cleaved PARP and cleaved caspase-9/8/3. These findings suggest that AETC induced NSCLC cell apoptosis via the ATF3-Hippo-YAP pathway in vivo and in vitro. We also found that AETC is non-toxic to normal cells and nude mice. Thus, AETC might represent a promising adjuvant for anti-tumor therapy against NSCLC.

Determining Pharmacological Mechanisms of Chinese Incompatible Herbs Fuzi and Banxia in Chronic Obstructive Pulmonary Disease: A Systems Pharmacology-Based Study.

Aconiti Lateralis Radix Praeparata (Fuzi) and Pinelliae Rhizoma (Banxia) are among the 18 incompatible medications that are forbidden from use in one formulation. However, there is increasing evidence implying that this prohibition is not entirely correct. According to the theory of Chinese traditional medicine, they can be used for the treatment of chronic obstructive pulmonary disease (COPD). Thus, we analyzed the possible approaches for the treatment of COPD using network pharmacology. The active compounds of Fuzi and Banxia (FB) were collected, and their targets were identified. COPD-related targets were obtained by analyzing the differentially expressed genes between COPD patients and healthy individuals, which were expressed using a Venn diagram of COPD and FB. Protein-protein interaction data and network regarding COPD and drugs used were obtained. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis were conducted. The gene-pathway network was constructed to screen the key target genes. In total, 34 active compounds and 47 targets of FB were identified; moreover, 7,153 differentially expressed genes were identified between COPD patients and healthy individuals. The functional annotations of target genes were found to be related to mechanisms such as transcription, cytosol, and protein binding; furthermore, 68 pathways including neuroactive ligand-receptor interaction, Kaposi sarcoma-associated herpesvirus infection, apoptosis, and measles were significantly enriched. FOS CASP3, VEGFA, ESR1, and PTGS2 were the core genes in the gene-pathway network of FB for the treatment of COPD. Our results indicated that the effect of FB against COPD may involve the regulation of immunological function through several specific biological processes and their corresponding pathways. This study demonstrates the application of network pharmacology in evaluating mechanisms of action and molecular targets of herb-opponents FB.

LncRNA MIR4435-2HG targets desmoplakin and promotes growth and metastasis of gastric cancer by activating Wnt/ß-catenin signaling.

Long non-coding RNAs (lncRNAs) have been implicated in the pathogenesis of gastric cancer; however, their mechanisms of action remain largely unknown. The aim of this study was to identify lncRNAs involved in the tumorigenesis of gastric cancer and to investigate the signaling pathways they affect. Using microarray and RT-qPCR analyses, candidate lncRNAs were screened in paired gastric cancer tissues. The analysis revealed MIR4435-2HG to be markedly up-regulated in gastric cancer samples compared to normal stomach specimens. Increased MIR4435-2HG expression was associated with aggressive clinicopathologic features and unfavorable tumor stage. Functional experiments showed that MIR4435-2HG up-regulation enhanced gastric cancer cell proliferation, clonogenicity, and migration and invasion in vitro, as well as tumorigenicity in mice. Using RNA pull-down and mass-spectrometry analyses we found and verified a direct and novel interaction between MIR4435-2HG and desmoplakin (DSP), the most abundant desmosomal protein. Overexpression and knockdown experiments revealed opposing roles for DSP and MIR4435-2HG, unmasking a cascade through which MIR4435-2HG binds to and inhibits DSP, leading to activation of WNT/ß-catenin signaling and epithelial-mesenchymal transition in gastric cancer cells. We propose that the MIR4435-2HG/DSP/WNT axis serves as a critical effector of carcinogenesis and progression of gastric cancer, and could be exploited therapeutically to improve patients' outcomes.

Role of Gut Microbiota in the Development and Treatment of Colorectal Cancer.

Human guts harbor abundant microbes that regulate many aspects of host physiology. However, bacterial imbalance or dysbiosis in the gut due to the dietary or environmental changes may cause colorectal cancer (CRC). Increasing studies show that gut microbiota plays an important role in the occurrence and development of CRC, as a result of virulence factors, bacterial metabolites, or inflammatory pathways. In the future, probiotics or targeting the microbiota will probably be a powerful weapon in the battle against CRC. This review seeks to outline the relationship between gut microbiota and the development of CRC as well as the potential mechanisms of microbiota involved in treatment of CRC, so as to provide some references for research on the development, prevention, and treatment of this disease.

Emergency obstetric hysterectomy for life-threatening postpartum hemorrhage: A 12-year review.

The aim of the study was to review the operative experiences of emergency hysterectomy for life-threatening postpartum hemorrhage (PPH) performed over a 12-year period at Fujian Provincial Maternity and Children's Hospital; to examine the incidence and risk factors for emergency obstetric hysterectomy; and to evaluate the curative effectiveness and safety of subtotal hysterectomy for life-threatening PPH.The records of all cases of emergency obstetric hysterectomy performed at Fujian Maternity and Children Health Hospital between January 2004 and June 2016 were analyzed. The incidence, risk factors, and outcomes of hysterectomy, the peripartum complications, and the coagulation function indices were evaluated.A total of 152,023 of women were delivered. The incidence of emergency postpartum hysterectomy was 0.63 per 1000 deliveries: 96 patients underwent hysterectomy for uncontrolled PPH, 19 (0.207‰) underwent hysterectomy following vaginal delivery, and 77 (1.28‰) underwent the procedure following cesarean delivery (P < .001). Common risk factors included postpartum prothrombin activity ≤ 50% (61.5%), placenta accreta (43.76%), uterine atony (37.5%), uterine rupture (17.5%), and grand multiparity > 6 (32.3%). Forty-one patients underwent subtotal abdominal hysterectomy (STH) and 55 patients underwent total abdominal hysterectomy (TH). The mean operation time was significantly shorter for TH (193.59 ±â€Š83.41 minutes) than for STH (142.86 ±â€Š78.32 minutes; P = .002). The mean blood loss was significantly greater for TH (6832 ±â€Š787 mL) than for STH (6329 ±â€Š893 mL; P = .003). The mean number of red cell units transfusion was higher during TH (16.24 ±â€Š9.48 units vs 12.43 ±â€Š7.2, respectively; P = .047). Postoperative prothrombin activity was significantly higher than preoperative levels (56.84 ±â€Š14.74 vs 44.39 ±â€Š15.69, respectively; P < .001) in women who underwent TH and in those who underwent STH (57.63 ±â€Š15.68 vs 47.87 ±â€Š12.86, respectively; P < .001). There was no significant difference in the maternal complications after TH or STH for PPH.Cesarean deliveries were associated with an increased risk of emergency hysterectomy, and postpartum prothrombin activity < 50% was the greatest risk factor for hysterectomy in most women who underwent hysterectomy. STH was the preferred procedure for emergency obstetric hysterectomy.

Analysis of intact and dissected fungal polyketide synthase-nonribosomal peptide synthetase in vitro and in Saccharomyces cerevisiae.

The widely found fungal iterative PKS-NRPS hybrid megasynthetases are highly programmed biosynthetic machines involved in the synthesis of 3-acyltetramic acids and related natural products. In vitro analysis of iterative PKS-NRPS has been hampered by the difficulties associated with obtaining pure and functional forms of these large enzymes (>400 kDa). We successfully expressed Aspergillus nidulans aspyridone synthetase (ApdA) from an engineered Saccharomyces cerevisiae strain. The complete functions of ApdA and its enoylreductase partner ApdC are reconstituted in vitro and in S. cerevisiae with the production of preaspyridone 7. The programming rules of both the PKS and NRPS modules were then examined in vitro. The key interaction between the PKS and the NRPS was dissected and reconstituted in trans by using stand-alone modules. Analogs of 7 were synthesized through heterologous combinations of PKS and NRPS modules from different sources. Our results represent one of the largest, multidomain enzyme reconstituted to date and offer new opportunities for engineered biosynthesis of fungal natural products.