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Abstract Purpose 46,XY disorders of sex development (46,XY DSD) are characterized by incomplete masculinization of genitalia with reduced androgenization. Accurate clinical management remains challenging, especially based solely on physical examination. Targeted next-generation sequencing (NGS) with known pathogenic genes provides a powerful tool for diagnosis efficiency. This study aims to identify the prevalent genetic variants by targeted NGS technology and investigate the diagnostic rate in a large cohort of 46,XY DSD patients, with most of them presenting atypical phenotypes. Methods Two different DSD panels were developed for sequencing purposes, targeting a cohort of 402 patients diagnosed with 46,XY DSD, who were recruited from the Department of Urology at Children’s Hospital, Zhejiang University School of Medicine (Hangzhou, China). The detailed clinical characteristics were evaluated, and peripheral blood was collected for targeted panels to find the patients’ variants. The clinical significance of these variants was annotated according to American College of Medical Genetics and Genomics (ACMG) guidelines. Results A total of 108 variants across 42 genes were found in 107 patients, including 46 pathogenic or likely pathogenic variants, with 45.7%(21/46) being novel. Among these genes, SRD5A2, AR, FGFR1, LHCGR, NR5A1, CHD7 were the most frequently observed. Besides, we also detected some uncommon causative genes like SOS1, and GNAS. Oligogenic variants were also identified in 9 patients, including several combinations PROKR2/FGFR1/CYP11B1, PROKR2/ATRX, PROKR2/AR, FGFR1/LHCGR/POR, FGFR1/NR5A1, GATA4/NR5A1, WNT4/AR, MAP3K1/FOXL2, WNT4/AR, and SOS1/FOXL2. Conclusion The overall genetic diagnostic rate was 11.2%(45/402), with an additional 15.4% (62/402) having variants of uncertain significance. Additionally, trio/duo patients had a higher genetic diagnostic rate (13.4%) compared to singletons (8.6%), with a higher proportion of singletons (15.1%) presenting variants of uncertain significance. In conclusion, targeted gene panels identified pathogenic variants in a Chinese 46,XY DSD cohort, expanding the genetic understanding and providing evidence for known pathogenic genes’ involvement.

Abstract Backgroud Type II congenital pulmonary airway malformation (CPAM) is a rare pulmonary microcystic developmental malformation. Surgical excision is the primary treatment for CPAM, although maternal steroids and betamethasone have proven effective in reducing microcystic CPAM. Disturbed intercellular communication may contribute to the development of CPAM. This study aims to investigate the expression profile and analyze intercellular communication networks to identify genes potentially associated with type II CPAM pathogenesis and therapeutic targets. Methods RNA sequencing (RNA-seq) was performed on samples extracted from both the cystic area and the adjacent normal tissue post-surgery in CPAM patients. Iterative weighted gene correlation network analysis (iWGCNA) was used to identify genes specifically expressed in type II CPAM. Single-cell RNA-seq (scRNA-seq) was integrated to unveil the heterogeneity in cell populations and analyze the communication and interaction within epithelial cell sub-populations. Results A total of 2,618 differentially expressed genes were identified, primarily enriched in cilium-related biological process and inflammatory response process. Key genes such as EDN1, GPR17, FPR2, and CHRM1, involved in the G protein-coupled receptor (GPCR) signaling pathway and playing roles in cell differentiation, apoptosis, calcium homeostasis, and the immune response, were highlighted based on the protein-protein interaction network. Type II CPAM-associated modules, including ciliary function-related genes, were identified using iWGCNA. By integrating scRNA-seq data, AGR3 (related to calcium homeostasis) and SLC11A1 (immune related) were identified as the only two differently expressed genes in epithelial cells of CPAM. Cell communication analysis revealed that alveolar type 1 (AT1) and alveolar type 2 (AT2) cells were the predominant communication cells for outgoing and incoming signals in epithelial cells. The ligands and receptors between epithelial cell subtypes included COLLAGEN genes enriched in PI3K-AKT singaling and involved in epithelial to mesenchymal transition. Conclusions In summary, by integrating bulk RNA-seq data of type II CPAM with scRNA-seq data, the gene expression profile and critical signaling pathways such as GPCR signaling and PI3K-AKT signaling pathways were revealed. Abnormally expressed genes in these pathways may disrupt epithelial-mesenchymal transition and contribute to the development of CPAM. Given the effectiveness of prenatal treatments of microcystic CPAM using maternal steroids and maternal betamethasone administration, targeting the genes and signaling pathways involved in the development of CPAM presents a promising therapeutic strategy.

Rapid urbanization and unplanned development have posed a threat to the thermal environment in a country like China. The urban heat island (UHI) phenomenon is one of the most serious issues because of its strong relation to thermal comfort, air pollution, and public health. The water bodies, as an important component of the urban ecosystem, are generally considered a vital resource to mitigate the UHI. This study provides direct evidence with the help of satellite observation and field measurement data using the mono-window algorithm, spatial buffer analysis, and linear regression methods to explore how the stream water affects the local temperature variation. The results showed that Land use/cover change (LUCC) and Land surface temperature (LST) on both banks of the Bahe River changed significantly. The LST of the river was significantly massively reduced within the distance range of 300 to 500 m, and 400 to 600 m, with an average temperature dip from 3.7 to 2.8 °C, and from 3.2 to 1.7 °C respectively were during the summer on east and west river banks. In addition, the surrounding LUCC composition and configuration could strongly affect the maximum cooling scale. The results provide insights for urban planners and managers to arrange the LUCC composition between the river design in urban areas and the cooling effect demands.

A broadband radar cross-section (RCS) reduction employed by the vortex metasurface is proposed. The underlying physics is associated with the Pancharatnam-Berry geometrical phases involved in spin-to-orbital angular momentum conversion. The proposed metasurface generates spin-controlled vortex waves with different topological charges. The vortex singularity implements specular extremely low RCS reduction. The prototypes are fabricated and measured to verify the wideband performance of the RCS reduction. The simulated results illustrate that −10 dB broadband RCS reduction from 9 GHz to 22.5 GHz and the RCS reduction has peak values −37.2 dB, −40.8 dB, and −42.5 dB at 9.8 GHz, 13.1 GHz, and 20.8 GHz, respectively. The measurement results illustrate that −10 dB RCS reduction over 9.1–18 GHz is realized, and the RCS reduction peak −53 dB at 10.1 GHz. The measured electric field intensities and phase distribution of the vortex metasurface also be displayed. Due to the limitation of experimental conditions, no test was done in the range greater than 18 GHz. Far-field results achieve good agreement between simulations and experiments, and the near-field measurements are a subtle difference in the experimental results. This work reveals the potential advantages of vortex waves in the fields of multifunctional electromagnetic stealth, beamforming for communication systems, etc.

Abstract Background Congenital pulmonary airway malformation (CPAM) is the most frequent pulmonary developmental malformation and the pathophysiology remains poorly understood. This study aimed to identify the characteristic gene expression patterns and the marker genes essential to CPAM. Methods Tissues from the cystic area displaying CPAM and the area of normal appearance were obtained during surgery. Bulk RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) were performed for integrating analysis. Iterative weighted gene correlation network analysis (iWGCNA) was used to identify specifically expressed genes to CPAM. Results In total, 2074 genes were significantly differentially expressed between the CPAM and control areas. Of these differentially expressed genes (DEGs), 1675 genes were up-regulated and 399 genes were down-regulated. Gene ontology analysis revealed these DEGs were specifically enriched in ciliated epithelium and involved in immune response. We also identified several CPAM-related modules by iWGCNA, among them, P15_I4_M3 module was the most influential module for distinguishing CPAMs from controls. By combining the analysis of the expression dataset from RNA-seq and scRNA-seq, SPOCK2, STX11, and ZNF331 were highlighted in CPAM. Conclusions Through our analysis of expression datasets from both scRNA-seq and bulk RNA-seq of tissues obtained from patients with CPAM, we identified the characteristic gene expression patterns associated with the condition. Our findings suggest that SPOCK2 could be a potential biomarker gene for the diagnosis and therapeutic target in the development of CPAM, whereas STX11 and ZNF331 might serve as prognostic markers for this condition. Further investigations with larger samples and function studies are necessary to confirm the involvement of these genes in CPAM.

Genetically abnormal fibroblasts are notably more prevalent in colorectal cancer (CRC) than in adjacent normal tissue, emphasizing their significance in driving the heterogeneity of the tumor microenvironment. Functioning as a significant regulatory gene in the context of fibrosis, FOXF1 adjacent non-coding developmental regulatory RNA (FENDRR) has exhibited abnormal expression in colorectal cancer and interstitial localization in our experiments. However, current research on the role of FENDRR in cancer has focused solely on its impact on cancer cells. Its crucial role in the tumor stroma is yet to be explored. The goal of this study was to understand the relationship between atypical FENDRR expression, its distinct localization, and genetically abnormal fibroblasts in CRC. We aimed to establish the function of FENDRR within the stromal compartment of patients through bioinformatics. Our study confirmed that FENDRR suppresses cancer-associated fibroblasts by inhibiting their activation and collagen generation in CRC. Furthermore, our findings suggest that low FENDRR expression indicates a poor prognosis. Therefore, we propose that FENDRR is a promising therapeutic target for future studies in CRC.

Abstract Sparganium longifolium was reported as a hybrid between S. emersum and S. gramineum based on its intermediate type or the common characteristics of its parent species. Its hybrid origin needs to be confirmed using molecular technology. We investigated the origin of S. longifolium based on 10 populations of S. emersum, S. gramineum and S. longifolium from five lakes in European Russia, using sequences of six nuclear loci and one chloroplast DNA fragment. Haplotype network, principal coordinate analysis and genetic clustering based on data of nuclear loci confirmed that S. longifolium is the hybrid between S. emersum and S. gramineum. We found that the natural hybridization between S. emersum and S. gramineum is bidirectional but asymmetrical, and the latter mainly acts as maternal species. We also found that all samples of S. longifolium were F1 generations, and thus hypothesized that S. emersum and S. gramineum could likely maintain their species boundary through the post-zygote reproductive isolation mechanism of F1 generation sterility.

Abstract Glomerular endothelial cells (GEnCs) dysfunction occurs at the early stage of diabetic nephropathy (DN). One of its characteristics is endothelial-to-mesenchymal transition (EndMT). Heparanase (HPSE) is the only known mammalian endoglycosidase capable of degrading heparin sulfates and has a prominent role in DN pathogenesis. However, whether HPSE induces EndMT of GEnCs remains unknown. This study aimed to determine the effect and potential mechanism of HPSE on GEnCs phenotype under high-glucose conditions. In the early development of streptozotocin (STZ)-induced diabetic mice, HPSE overexpression was positively correlated with renal injury and the number of GEnCs undergoing EndMT, which was characterized by loss of endothelial marker CD31 and gain of mesenchymal markers including α-SMA and Snail1/2 by double immunofluorescence staining. Bioinformatics analysis revealed a positive correlation between HPSE and ERK. The counts of double positive staining of CD31 and p-ERK1/2 was significantly increased in the glomeruli of STZ-induced diabetic mice compared with sham mice. In cultured GEnCs, high glucose dramatically upregulated the expressions of HPSE and p-ERK1/2, both of which were markedly blocked by HPSE siRNA. Furthermore, recombinant mouse HPSE (rmHPSE) promoted the expressions of mesenchymal markers and p-ERK1/2 in a dosage- and time-dependent manner. U0126, a specific MEK/ERK inhibitor, significantly inhibited either high glucose or rmHPSE-induced EndMT of GEnCs. These data indicate that high glucose induces EndMT of GEnCs at least partially through upregulating HPSE and that HPSE promotes EndMT of GEnCs via activating ERK signaling. This study improves understanding the crucial role of HPSE in DN development and progression.

Abstract Background Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest malignancies to date. The impressively developed stroma that surrounds and modulates the behavior of cancer cells is one of the main factors regulating the PDAC growth, metastasis and therapy resistance. Here, we postulate that stromal and cancer cell compartments differentiate in protein/lipid glycosylation patterns and analyze differences in glycan fragments in those compartments with clinicopathologic correlates. Results We analyzed native glycan fragments in 109 human FFPE PDAC samples using high mass resolution matrix-assisted laser desorption/ionization Fourier-transform ion cyclotron resonance mass spectrometric imaging (MALDI-FT-ICR-MSI). Our method allows detection of native glycan fragments without previous digestion with PNGase or any other biochemical reaction. With this method, 8 and 18 native glycans were identified as uniquely expressed in only stromal or only cancer cell compartment, respectively. Kaplan–Meier survival model identified glycan fragments that are expressed in cancer cell or stromal compartment and significantly associated with patient outcome. Among cancer cell region-specific glycans, 10 predicted better and 6 worse patient survival. In the stroma, 1 glycan predicted good and 4 poor patient survival. Using factor analysis as a dimension reduction method, we were able to group the identified glycans in 2 factors. Multivariate analysis revealed that these factors can be used as independent survival prognostic elements with regard to the established Union for International Cancer Control (UICC) classification both in tumor and stroma regions. Conclusion Our method allows in situ detection of naturally occurring glycans in FFPE samples of human PDAC tissue and highlights the differences among glycans found in stromal and cancer cell compartment offering a basis for further exploration on the role of specific glycans in cancer–stroma communication.

AbstractTo investigate the fracture propagation mechanism during supercritical CO2 fracturing in shale reservoirs, a numerical model was proposed based on the displacement discontinuity method. The Peng–Robinson equation was introduced to determine the variations in CO2 properties during the fracturing process. Considering natural fracture distribution in shale reservoirs, the fracture propagation mechanisms during supercritical CO2 fracturing in shale reservoirs under different horizontal stress differences and matrix permeabilities were analyzed. The influence of the proportion of CO2 preenergizing on fracture morphology was discussed. The results obtained via numerical simulation show that supercritical CO2 is beneficial to create a more complex fracture network by activating natural fractures under the same horizontal stress difference. CO2 easily penetrates into the matrix near the fracture surfaces, increasing reservoir energy. However, when the permeability of shale reservoirs exceeds 0.04×10−3 μm2, substantial filtration of CO2 into the reservoir matrix occurs near the well bore, limiting the activation of natural fractures around the fracture tip. A higher proportion of CO2 preenergizing during fracturing is conducive to improve the fracture complexity while reducing the fracture aperture.

Background: Obesity is a principal risk factor for the development of type 2 diabetes and cardiovascular diseases. Natural plants and/or foods play an important role in the management of obesity. Acalypha australis L. (AAL) is a kind of potherb popular among Asian populations, and it is also consumed as a food ingredient and traditional herbal medicine. Objective: We investigated the effects of water extract from AAL on high-fat-diet (HFD)-induced obese mice and 3T3-L1 adipocytes to develop a new functional food material. Design: Nine-week-old male mice were randomly divided into control (chow diet, n = 6) and HFD (n = 30) group. From 12-weeks onward, mice in the HFD group were further separated into model (saline, 6 mL/kg), simvastatin (0.11 mg/mL, 6 mL/kg), and AAL treatment (low, middle, and high dosage: 300, 600, and 900 mg/kg) group, with 6 animals per group, while mice in the control group were treated with saline (6 mL/kg). Food intake, body/fat weight, liver/kidney indexes, and lipid profiles were determined. Tissues were fixed with formalin for pathological examination. Western blotting and PCR were performed to evaluate the protein and mRNA expression in 3T3-L1 adipocytes. Oil Red O staining was used to determine lipid accumulation. Results: AAL administration significantly suppressed body weight gain, and reduced fat pad weight and Lee’s index in obese mice, but had no effect on liver/kidney index. AAL also reduced serum cholesterol, triglyceride, and LDL-C and increased HDL-C levels. Histological analysis revealed that AAL significantly ameliorated lipid accumulation in the liver and subcutaneous adipose tissue. In vitro, Oil Red O staining showed that AAL inhibited adipose differentiation by down-regulating the gene and protein expression of PPARγ and C/EBPα. AAL also reversed HFD-induced intestinal dysbacteriosis. Conclusion: AAL water-soluble extract has a significant anti-adipogenic effect in the HFD-induced obese mice model.

Abstract Background Urinary retention (UR) is a common urinary system disease can be caused by urinary tract obstruction with numerous reasons, however, the role of urine microbes in these disorders is still poorly understood. The aim of this study was to identify the urine microbial features of two common types of obstructive UR, caused by urinary stones or urinary tract tumors, with comparison to healthy controls. Methods Urine samples were collected from a cohort of 32 individuals with stone UR, 25 subjects with tumor UR and 25 healthy controls. The urine microbiome of all samples was analyzed using high-throughput 16S rRNA (16S ribosomal RNA) gene sequencing. Results We observed dramatically increased urine microbial richness and diversity in both obstructive UR groups compared to healthy controls. Despite different origins of UR, bacteria such as Pseudomonas, Acinetobacter and Sphingomonas were enriched, while Lactobacillus, Streptococcus, Gardnerella, Prevotella and Atopobium were decreased in both UR groups in comparison with healthy controls, exhibited an approximate urine microbial community and functional characteristics of two types of obstructive UR. Furthermore, disease classifiers were constructed using specific enriched genera in UR, which can distinguish stone UR or tumor UR patients from healthy controls with an accuracy of 92.29% and 97.96%, respectively. Conclusion We presented comprehensive microbial landscapes of two common types of obstructive urinary retention and demonstrated that urine microbial features of these patients are significantly different from that of healthy people. The urine microbial signatures would shed light on the pathogenesis of these types of urinary retention and might be used as potential classification tools in the future.

Abstract Expanded newborn screening facilitates early identification and intervention of patients with inborn errors of metabolism (IEMs), There is a lack of disease spectrum data for many areas in China. To determine the disease spectrum and genetic characteristics of IEMs in Xi'an city of Shaanxi province in northwest China, 146152 newborns were screening by MSMS from January 2014 to December 2019 and 61 patients were referred to genetic analysis by next generation sequencing (NGS) and validated by Sanger sequencing. Seventy-five newborns and two mothers were diagnosed with IEMs, with an overall incidence of 1:1898 (1:1949 without mothers). There were 35 newborns with amino acidemias (45.45%, 1:4176), 28 newborns with organic acidurias (36.36%, 1:5220), and 12 newborns and two mothers with FAO disorders (18.18%; 1:10439 or 1:12179 without mothers). Phenylketonuria and methylmalonic acidemia were the two most common disorders, accounting for 65.33% (49/75) of all confirmed newborn. Some hotspot mutations were observed for several IEMs, including PAH gene c.728G>A for phenylketonuria; MMACHC gene c.609G>A and c.567dupT, MMUT gene c.323G>A for methylmalonic acidemia and SLC25A13 gene c.852_855del for citrin deficiency. Our study provides effective clinical guidance for the popularization and application of expanded newborn screening, genetic screening, and genetic counseling of IEMs in this region.

The main area of the Jiaoshiba anticline of the Fuling shale gas field was taken as the research object, laboratory rock mechanical experiments and direct shear experiments were conducted to clarify the mechanical anisotropy characteristics and parameters of rock samples with rich beddings. Based on the experimental results, a 3D fracture propagation model of the target reservoir taking mechanical anisotropy, weak bedding plane and vertical stress difference into account was established by the discrete element method to analyze distribution patterns of hydraulic fractures under different bedding densities, mechanical properties, and fracturing engineering parameters (including perforation clusters, injection rates and fracturing fluid viscosity). The research results show that considering the influence of the weak bedding plane and longitudinal stress difference, the interlayer stress difference 3–4 MPa in the study area can control the fracture height within the zone of stress barrier, and the fracture height is less than 40 m. If the influence of the weak bedding plane is not considered, the simulation result of fracture height is obviously higher. Although the opening of high-density bedding fractures increases the complexity of hydraulic fractures, it significantly limited the propagation of fracture height. By reducing the number of clusters, increasing the injection rate, and increasing the volume and proportion of high-viscosity fracturing fluid in the pad stage, the restriction on fracture height due to the bedding plane and vertical stress difference can be reduced, and the longitudinal propagation of fractures can be promoted. The fracture propagation model was used to simulate one stage of Well A in Fuling shale gas field, and the simulation results were consistent with the micro-seismic monitoring results.

Ball-sealer in-stage diversion in horizontal wells is a key technique to realize the uniform stimulation of fractured sections in tight oil and gas reservoirs. So far, however, there are fewer research results on the propagation morphologies of multi-cluster fractures after the implementation of different ball-sealer in-stage diversion processes during the fracturing treatment, which results in less theoretical support for the preparation of field ball-sealer in-stage diversion process and measures and impacts its application effects in the fracturing field. To deal with this situation, this paper established a fully coupled “wellbore–perforation–fracture propagation” model of horizontal wells on the basis of the boundary element method. Then, a method for calculating the allocation of ball sealers was proposed. Finally, the number of ball sealers, diversion time and number of diversions during the intra-stage temporary plugging and diversion under the condition of initial heterogeneous stress field and their effects on the propagation of multi-cluster fractures were simulated. And the following research results were obtained. First, the flow restriction of perforation friction can counterbalance the intake difference caused by the induced stress interference so that the friction difference applied on the fluid flow in each fracture cluster is reduced. Second, when the effect of the heterogeneous distribution of the initial stress field is taken into consideration, the liquid intake of each fracture cluster changes greatly and even ineffective perforation clusters without liquid incoming appear in the high-stress region. And after the ball is injected, new fractures are initiated from the ineffective perforation clusters. Third, when the initial minimum horizontal principal stress difference (Δσh) is higher than 3 MPa, it is beneficial to reduce the non-uniform propagation of each fracture cluster by increasing the number of ball sealers appropriately in the middle stage of the construction (over half of the total perforations of each stage) or carrying out temporary plugging in the early–middle stage (including ball injection in batches in the early–middle stage). Fourth, when Δσh is lower than 2 MPa, it is necessary to reduce the number of ball sealers or inject balls in the middle–late stage, or the non-uniform propagation of each fracture cluster will be aggravated.

Abstract Tobacco (Nicotiana tabacum) is considered as the model plant for alkaloid research, of which nicotine accounts for 90%. Many nicotine biosynthetic genes have been identified and were known to be regulated by jasmonate-responsive transcription factors. As an important regulator in plant physiological processes, whether small RNAs are involved in nicotine biosynthesis is largely unknown. Here, we combine transcriptome, small RNAs and degradome analysis of two native tobacco germplasms YJ1 and ZY100 to investigate small RNA’s function. YJ1 leaves accumulate twofold higher nicotine than ZY100. Transcriptome analysis revealed 3,865 genes which were differently expressed in leaf and root of two germplasms, including some known nicotine and jasmonate pathway genes. By small RNA sequencing, 193 miRNAs were identified to be differentially expressed between YJ1 and ZY100. Using in silico and degradome sequencing approaches, six nicotine biosynthetic genes and seven jasmonate pathway genes were predicted to be targeted by 77 miRNA loci. Three pairs among them were validated by transient expression in vivo. Combined analysis of degradome and transcriptome datasets revealed 51 novel miRNA-mRNA interactions that may regulate nicotine biosynthesis. The comprehensive analysis of our study may provide new insights into the regulatory network of nicotine biosynthesis.

CO2-based fracturing is widely introduced to stimulate shale oil reservoirs for its multiple advantages. However, the range of CO2 entering the matrix around fractures and CO2-oil replacement capacity between matrix and fractures cannot be fully explained. To address this issue, a radial constant volume diffusion experiment on shale cores was designed in this study, and the pressure drop curve history was matched through numerical model to determine the composition effective diffusion coefficient. A field-scale numerical model was established, in which a series of certain grids were used to explicitly characterize fracture and quantify the prosess of CO2 mass transfer and oil replacement. Based on the field-scale numerical model, the process of shut-in, flow back, and oil production was simulated. The distribution of CO2 in fractured shale oil formation and its impact on crude oil during shut-in stage and flow back stage were investigated. This study concludes that CO2 gradually exchanges the oil in matrix into fractures and improve the fluidity of oil in matrix until the component concentrations of the whole reservoir reaches equilibrium during the shut-in process. Finally, about 30∼35 mole % of CO2 in fractures exchanges for oil in matrix. The range of CO2 entering the matrix around fractures is only 1.5 m, and oil in matrix beyond this distance will not be affected by CO2. During the process of flow back and production, the CO2 in fracture flows back quickly, but the CO2 in matrix is keeping dissolved in oil and will not be quickly produced. It is conclued that the longest possible shut-in time is conducive to making full use of the CO2-EOR mechanism in fractured shale oil reservoirs. However, due to the pursuit of economic value, a shut-in time of 10 days is the more suitable choice. This work can provide a better understanding of CO2 mass transfer mechanism in fractured shale oil reservoirs. It also provides a reference for the evaluation of the shut-in time and production management after CO2 fracturing.

Insulin-like growth factor-1 (IGF-1) function declines with age and is associated with brain ageing and the progression of age-related neurological conditions. The reversible binding of IGF-1 to IGF binding protein (IGFBP)-3 regulates the amount of bioavailable, functional IGF-1 in circulation. Cyclic glycine-proline (cGP), a metabolite from the binding site of IGF-1, retains its affinity for IGFBP-3 and competes against IGF-1 for IGFBP-3 binding. Thus, cGP and IGFBP-3 collectively regulate the bioavailability of IGF-1. The molar ratio of cGP/IGF-1 represents the amount of bioavailable and functional IGF-1 in circulation. The cGP/IGF-1 molar ratio is low in patients with age-related conditions, including hypertension, stroke, and neurological disorders with cognitive impairment. Stroke patients with a higher cGP/IGF-1 molar ratio have more favourable clinical outcomes. The elderly with more cGP have better memory retention. An increase in the cGP/IGF-1 molar ratio with age is associated with normal cognition, whereas a decrease in this ratio with age is associated with dementia in Parkinson disease. In addition, cGP administration reduces systolic blood pressure, improves memory, and aids in stroke recovery. These clinical and experimental observations demonstrate the role of cGP in regulating IGF-1 function and its potential clinical applications in age-related brain diseases as a plasma biomarker for—and an intervention to improve—IGF-1 function.

AbstractShale gas reservoirs have gradually become the main source for oil and gas production. The automatic optimization technology of complex fracture network in fractured horizontal wells is the key technology to realize the efficient development of shale gas reservoirs. In this paper, based on the flow model of shale gas reservoirs, the porosity/permeability of the matrix system and natural fracture system is characterized. The fracture network morphology is finely characterized by the fracture network expansion calculation method, and the flow model was proposed and solved. On this basis, the influence of matrix permeability, matrix porosity, fracture permeability, fracture porosity, and fracture length on the production of shale gas reservoirs is studied. The optimal design of fracture length and fracture location was carried, and the automatic optimization method of complex fracture network parameters based on simultaneous perturbation stochastic approximation (SPSA) was proposed. The method was applied in a shale gas reservoir, and the results showed that the proposed automatic optimization method of the complex fracture network in shale gas reservoirs can automatically optimize the parameters such as fracture location and fracture length and obtain the optimal fracture network distribution matching with geological conditions.

BackgroundChronic liver fibrosis is an inevitable stage for the development of patients with chronic hepatitis B (CHB). However, anti-fibrotic therapies have been unsuccessful so far. The biological functions and molecular mechanisms of long non-coding RNAs (lncRNAs) in the host immune system during chronic hepatitis B virus (HBV) infection, especially in fibrosis, are still largely unknown.MethodThe total RNA of peripheral blood mononuclear cells (PBMCs) from asymptomatic carriers (ASCs) or CHB receiving at least 8 years of anti-viral treatments was analyzed using Arraystar microarray and validated via quantitative real-time PCR (qRT-PCR). Correlation analysis was conducted based on correlation coefficients, Clusterprofile, and RNA Interactome Database (RAID). The functions of lncRNA in monocytes were determined via loss-of-function RNAi or gain-of-function lentivirus assays. The expression levels of mRNAs or proteins were evaluated using qRT-PCR, western blotting assay, or enzyme linked immunosorbent assays (ELISA).ResultsA total of 1,042 mRNA transcripts (630 up-regulated and 412 down-regulated) were identified being differentially expressed between ASC and CHB patients. Through enrichment analysis we focused on the transforming growth factor beta (TGF-β) signaling pathway and validated their expression in a larger cohort. Moreover, we found that lncRNA ENST00000519726 (lncRNA-HEIM) was highly expressed in monocytes and further up-regulated upon HBV infection. LncRNA-HEIM played an important role in CHB patients with long-term antiviral treatments, and its elevated expression was remarkably correlated with the TGF-β signaling pathway, especially with the two members namely TGF-β and SMAD4. Furthermore, altering the endogenous lncRNA-HEIM level in monocytes significantly affected the production of TGF-β, as well as the fibrosis of hepatic stellate cells by affecting the expression of collagen I and α-smooth muscle actin (α-SMA).ConclusionThese findings not only added knowledge to the understanding of the roles of which lncRNA-HEIM played in the activation of HSCs in CHB patients with long-term medication, but also provided a promising therapeutic target in the future treatment for liver fibrosis.

We extend the array theory to design 2D curved coding metasurfaces in order to achieve arbitrary wavefront control. The extended array theory establishes the relationship between wavefront and curved-surface, which provides a simple and effective method to obtain the layout of desired metasurfaces. According the theory, we present 2D curved metasurfaces with 3-bit coding elements. Both simulations and measurements demonstrate that the proposed designs can manipulate wavefront effectively. For normal incidence, the aperture efficiency of normal reflection is more than 90%. For general oblique incidence, the proposed designs almost meet the expectances, but the aperture efficiency decreases with the increase of desired scattering elevation angle. The curved coding metasurfaces are expected to be applied to many devices such as conformal antenna array.

The Dongsheng gas field is characterized by low porosity and low permeability. Its principal producing reservoir is the H-1 zone, belonging to the Lower Shihezi Formation. Sand is the main lithology in the H-1 zone, while mud interlayers are also well developed in a vertical direction. As a result, the reservoir is a sand-mud interbed, which brings difficulty to fracture height extension. In order to understand the process of fracture height growth in a sand-mud interbed reservoir and obtain the responses of injection pressure and fracture width during a hydraulic fracturing, a hydromechanical-coupled model is established. Mud interlayers are fully considered and a cohesive zone model is adopted to deal with fracture propagation. Numerical results show that the fracture extends quickly to the sand-mud interface after initiation and breaks through rather than propagating along the interfaces. Pressure and width both increase continuously when fracture propagates in the mud interlayer. High-viscosity and high-injection rates are helpful for the fracture to break through the mud interlayer, especially at an early period. When the mud interlayers are asymmetric, pressure and width fluctuate several times once fracture propagates inside and breaks through the mud interlayer. Perforations close to the thinner mud interlayer can increase the fracture width and reduce fracturing risks.

To describe the fracture width is crucial for the flow conductivity evaluation, which influences the exploitation efficiency of unconventional oil and gas resources. Commonly, as the proppants fill in the fracture, the deformation will happen under the closure pressure to resist the fracture width change. Therefore, it is significant to develop the theoretical model to predict the variation. In this work, the mathematical model for the propping behavior of proppants in the fracture under closure pressure is established based on the Hertz contact theory. Compared with the existing models, the developed model considers both the proppant insertion and the elastic compression among the proppants, which is closer to the actual physical process. Furthermore, the experimental cases with different proppant sizes are taken to verify the model, and the good conformity presents its rationality. The parameter sensitivity analysis of this model shows that the fracture width change increases with the increase of the average diameters of proppants (D) and it declines with the improving of proppant elasticity modulus (E1) and Poisson’s ratio (v1).

Nano secondary ion mass spectrometry (nanoSIMS) imaging is a rapidly growing field in biological sciences, which enables investigators to describe the chemical composition of cells and tissues with high resolution. One of the major challenges of nanoSIMS is to identify specific molecules or organelles, as these are not immediately recognizable in nanoSIMS and need to be revealed by SIMS-compatible probes. Few laboratories have generated such probes, and none are commercially available. To address this, we performed a systematic study of probes initially developed for electron microscopy. Relying on nanoscale SIMS, we found that antibodies coupled to 6 nm gold particles are surprisingly efficient in terms of labeling specificity while offering a reliable detection threshold. These tools enabled accurate visualization and sample analysis and were easily employed in correlating SIMS with other imaging approaches, such as fluorescence microscopy. We conclude that antibodies conjugated to moderately sized gold particles are promising tools for SIMS imaging.

The homeostasis of NAD+ anabolism is indispensable for maintaining the NAD+ pool. In mammals, the mainly synthetic pathway of NAD+ is the salvage synthesis, a reaction catalyzed by nicotinamide mononucleotide adenylyltransferase (NAMPT) and nicotinamide mononucleotide adenylyltransferase (NMNATs) successively, converting nicotinamide (NAM) to nicotinamide mononucleotide (NMN) and NMN to NAD+, respectively. However, the relationship between NAD+ anabolism disturbance and diabetic nephropathy (DN) remains elusive. Here our study found that the disruption of NAD+ anabolism homeostasis caused an elevation in both oxidative stress and fibronectin expression, along with a decrease in Sirt1 and an increase in both NF-κB P65 expression and acetylation, culminating in extracellular matrix deposition and globular fibrosis in DN. More importantly, through constitutively overexpressing NMNAT1 or NAMPT in human mesangial cells, we revealed NAD+ levels altered inversely with NMN levels in the context of DN and, further, their changes affect Sirt1/NF-κB P65, thus playing a crucial role in the pathogenesis of DN. Accordingly, FK866, a NAMPT inhibitor, and quercetin, a Sirt1 agonist, have favorable effects on the maintenance of NAD+ homeostasis and renal function in db/db mice. Collectively, our findings suggest that NMN accumulation may provide a causal link between NAD+ anabolism disturbance and diabetic nephropathy (DN) as well as a promising therapeutic target for DN treatment.

The shale gas exploration and development in Sichuan Basin had achieved a major breakthrough remarkably. However, the business development focused on the marine shale gas in shallow layers (vertical depth 3500 m) business development in Wufeng Formation-Longmaxi Formation is the key to the rapid growth of shale gas production in the Sichuan Basin. In the literature, authors analyzed the geological characteristics of the deep shale gas of the Wufeng Formation - Longmaxi Formation in southern Sichuan Basin through particular dissection and analysis of deep wells in comparison to the shallow layers. The thickness of organic-rich shale in the deep region of southern Sichuan Basin measures to be more than 120 m, which is larger than that of the shallow layers in Fuling area. The vertical TOC change remains constant whereas the thickness of high-quality shale is approximately forty meters in both regions. Nevertheless, the RO is higher in the deep region of southern Sichuan Basin. The calcareous content of the organic shale increases while the silicon content decreases from the Fuling area down as it gets deeper into the region in southern Sichuan Basin. It is quite evident in research that the shales in the Wufeng Formation – Longmaxi Formation of the deep to shallow layers of southern Sichuan that there are not many differences in depositional environment, lithology, TOC, etc. However, there are four major differences: (1) The porosity of shale in the deep region of southern Sichuan Basin is higher than that in Fulin gas field due to the corrosion pore. (2) The gas-bearing properties are better in the deep regions of southern Sichuan Basin because they have better preservation condition compared to that of the shallow region of Fuling. (3) However, gas-bearing properties are worse in the local region of stronger structure deformation and the basin boundary fault zone. (4) The shale gas resources in the deep regions of southern Sichuan Basin are great and have wide prospects for exploration and development. In order for a breakthrough to be generated in deep shale gas commercial development in the soonest possible time, authors propose the following: utilize strengthening evaluation of small layer contrasting so as to explain fine structure and predict fracture; strengthen the research on drilling and fracturing technology for deep shale gas; strengthen research on wells column; and support technology for efficient production of shale gas. Keywords: Sichuan basin, Lower Silurian, Longmaxi formation, Deep shale gas, Marine facies

Alterations to the gut microbiota have been previously suggested to be tightly linked to chronic systemic inflammation, which is a major contributing factor to complications and disease progression in chronic kidney disease (CKD). Nevertheless, the effect of gut dysbiosis on the pathogenesis and/or production of inflammatory factors in CKD has not been extensively studied to date. In the present study, we conducted 16S ribosomal DNA pyrosequencing using fecal microbiota samples and analyzed the production of serum inflammatory factors in 50 patients with CKD and 22 healthy control (HC) subjects. The results revealed that compared to the HC subjects, patients with CKD exhibited a significant reduction in the richness and structure of their fecal microbiota. At the phylum level, compared to the HC group, patients with CKD also presented reduced abundance of Actinobacteria but increased abundance of Verrucomicrobia. Moreover, the genera Lactobacillus, Clostridium IV, Paraprevotella, Clostridium sensu stricto, Desulfovibrio, and Alloprevotella were enriched in the fecal samples of patients with CKD, while Akkermansia and Parasutterella were enriched in those of the HC subjects. The abundance of Akkermansia in the CKD group was significantly lower than that in the HC group (3.08 vs. 0.67%); this decrease in the abundance of Akkermansia, an important probiotic, in patients with CKD is a striking discovery as it has not been previously reported. Finally, we analyzed whether these changes to the fecal microbiota correlated with CKD clinical characteristics and/or the production of known inflammatory factors. Altered levels of the microbiota genera Parasutterella, Lactobacillus, Paraprevotella, Clostridium sensu stricto, and Desulfovibrio were shown to be correlated with CKD disease-severity indicators, including the estimated glomerular filtration rate. Most notably, Akkermansia was significantly negatively correlated with the production of interleukin-10. The results of the present study suggest that microbiota dysbiosis may promote chronic systemic inflammation in CKD. Furthermore, they support that modifying the gut microbiota, especially Akkermansia, may be a promising potential therapeutic strategy to attenuate the progression of, and/or systemic inflammation in, CKD.

Primary Sjögren's syndrome (pSS) is a common chronic autoimmune disease characterized by a high prevalence of autoantibodies and lymphocyte-mediated exocrine gland damage. To enhance our understanding of the mechanisms underlying the progression of the disease and to discover potential biomarkers for the early diagnosis of pSS, we applied RNA sequencing to compare the gene expression patterns in minor salivary glands between pSS patients and non-pSS. A total of 293 differentially expressed genes (DEGs) were detected in pSS vs. non-pSS (FDR < 0.05, fold changes > 2). Of these DEGs, 285 (97.26%) were up-regulated, with most being involved in immune system activation, especially in the formation of the immunological synapse. Significantly elevated CCL19/CCR7 expression in the salivary gland was found to be related to anti-Sjögren's syndrome-related antigen A (SSA) antibody and IgG levels in pSS patients, which was further confirmed in a larger cohort. Up-regulated gene expression showed strong discriminatory accuracy in identifying pSS with area under the curve of 0.98 using receiver operating characteristic curve analysis. In conclusion, gene expression changes in pSS include strong markers of immunological activation and have good discriminatory power in identifying patients with pSS.

High levels of salinity induce serious oxidative damage in plants. Flavonoids, as antioxidants, have important roles in reactive oxygen species (ROS) scavenging. In the present study, the tobacco R2R3 MYB type repressor, NtMYB4, was isolated and characterized. The expression of NtMYB4 was suppressed by salinity. Overexpression of NtMYB4 reduced the salt tolerance in transgenic tobacco plants. NtMYB4 repressed the promoter activity of NtCHS1 and negatively regulated its expression. Rutin accumulation was significantly decreased in NtMYB4 overexpressing transgenic plants and NtCHS1 RNAi silenced transgenic plants. Moreover, high H2O2 and O2− contents were detected in both types of rutin-reduced transgenic plants under high salt stress. In addition, exogenous rutin supplementation effectively scavenged ROS (H2O2 and O2−) and improved the salt tolerance of the rutin-reduced transgenic plants. In contrast, NtCHS1 overexpressing plants had increased rutin accumulation, lower H2O2 and O2− contents, and higher tolerance to salinity. These results suggested that tobacco NtMYB4 acts as a salinity response repressor and negatively regulates NtCHS1 expression, which results in the reduced flavonoid accumulation and weakened ROS-scavenging ability under salt stress.

Hydraulic fracturing using freshwater is difficult for the commercial exploitation of a shale oil reservoir in Jianghan Basin with a continental saline lake basin sedimentary background. Supercritical CO2 (SC-CO2) is a promising fracturing fluid under consideration for the reservoir stimulation, especially in the case of the presence of water sensible salt layers. In this study, SC-CO2 fracturing experiments on the inter-salt-shale and salt specimens, which were obtained from the drilling well, were carried out in the laboratory. The characteristics of fracture propagation, including morphology and width variation, were analyzed based on the observations of a stereoscopic microscope, X-ray micro-CT scanner, and 3D scanner. The existing weak planes in the shale can really impact the fracture propagation in SC-CO2 fracturing. Deflection, branching, and approaching can occur during the process of fracture propagation. The average width value of the reactivated natural fracture is bigger than that of a newly created fracture. In addition, the fracturing results indicate the greater breakdown pressure of rock salt if compared with the inter-salt-shale. The induced fractures in the salt specimen are compact and smaller in average width than those in the shale specimen. The higher breakdown pressure and relatively smaller fracture width of rock salt are real challenges for the fracturing of an inter-salt-shale oil reservoir.

The formation of complex fracture networks through the fracturing technology is a crucial operation used to improve the production capacity of tight gas/oil. In this study, physical simulation experiments of hydraulic fracturing were conducted with a true triaxial test system on cubic shale oil samples from the Yanchang Formation, China. The fractures were scanned by CT both before and after the experiments and then reconstructed in 3D. The complexity of fracture networks was investigated quantitatively by the fractal theory with topology. Finally, the effect of the horizontal stress ratio, fluid viscosity, and natural fractures on the complexity of the fracture networks was discussed. The results indicate that the method based on fractal theory and topology can effectively characterize the complexity of the fracture network. The change rates of the fractal dimension (K) are 0.45–3.64%, and the fractal dimensions (DNH) of the 3D fracture network after fracturing are 1.9522–2.1837, the number of connections per branch after fracturing (CB) are 1.57–2.0. The change rate of the fractal dimension and the horizontal stress ratio are negatively correlated. However, the change rate of the fractal dimension first increases and then decreases under increasing fluid viscosities, and a transition occurs at a fluid viscosity of 5.0 mPa·s. Whether under different horizontal stress ratios or fluid viscosities, the complexity of the fracture networks after fracturing can be divided into four levels according to DNH and CB. Complex fracture networks are more easily formed under a lower horizontal stress ratio and a relatively low fluid viscosity. A fracturing fluid viscosity that is too low or too high limits the formation of a fracture network.

sua-CMS (cytoplasmic male sterility) is the only male sterile system in tobacco breeding, but the mechanism of abortion is unclear. Cytological characteristics show that abortion in the sua-CMS line msZY occurs before the differentiation of sporogenous cells. In this study, a comparative transcriptomic analysis was conducted on flower buds at the abortion stage of msZY and its male fertile control ZY. A total of 462 differentially expressed genes were identified in msZY and ZY, which were enriched via protein processing in the endoplasmic reticulum (ER), oxidative phosphorylation, photosynthesis, and circadian rhythm-plant by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Most genes were downregulated in the ER stress pathway, heat-shock protein family, F1F0-ATPase encoding by the mitochondrial genome, and differentiation of stamens. Genes in the programmed cell death (PCD) pathway were upregulated in msZY. The transcriptome results were consistent with those of qRT-PCR. Ultrastructural and physiological analyses indicted active vacuole PCD and low ATP content in msZY young flower buds. We speculated that PCD and a deficiency in ATP synthesis are essential for the abortion of sua-CMS. This study reveals the potential mechanism of abortion of tobacco sua-CMS.

Abstract Objective The purpose of this study is to identify and visualize from different perspectives the topic on occupational exposure in operating room (OEOR). Methods In the Web of Science Core Collection (WoSCC), all the half-century data were retrieved from January 1st, 1973 to December 31st, 2022. CiteSpace, VOSviewer and Excel 2019 were employed to analyze and visualize data, based on publications, countries, institutions, journals, authors, keywords. Result A total of 336 journal papers were found. The increase of publications virtually started in 1991, peaked in 2020 and has been slowing down ever since. USA played most significant part among all the 49 countries/regions, while Universidade Estadual Paulista out of 499 institutions published the most papers. International Archives of Occupational and Environmental Health bears the most documents and citations in all the 219 retrieved journals. There are 1847 authors found, among whom Hoerauf K is the most influential one. "Occupational exposure”, “nitrous oxide” and “operating room personnel” are the top 3 co-occurrences keywords. Conclusion The trend in the field lies in “anaesthetic gas”, “blood borne pathogen”, “radiation” and “aerosol”, while “surgical smoke” and “occupational safety” are the recently researching hot spots in this study. Accurate recognize and effective protection are always essential subjects for researchers.