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Rationale: A novel laser ablation‐isotope ratio mass spectrometry (LA‐IRMS) method for in situ analysis of sulfur isotopes in sulfides has been developed. Instead of the in situ reaction applied by the traditional laser microprobe, the analyte gas preparation in this method is separated temporally and spatially from the LA, resulting in improved precision and accuracy. Methods: Our LA‐IRMS system combines an ultraviolet LA system, an elemental analyzer (EA), a custom‐built cryogenic concentration system, a continuous‐flow interface, and an IRMS. The sulfide aerosol particles generated from LA were transferred by a helium carrier gas from the ablation cell into the reaction tube and were then converted into SO2. Subsequently, SO2 was enriched in two cold traps and was finally introduced into the ion source of the IRMS through the continuous‐flow interface. Results: We measured three synthetic and four natural sulfide reference materials to test the performance of this method. Precisions of ±0.25‰–±0.48‰ and ±0.32‰–±0.64‰ (1SD, n = 5) for δ34S values of synthetic and natural sulfide standards can be obtained for spot sizes ranging from 64 to 80 μm. Measured values and their recommended values showed a good linear relationship (R2 within 0.998 and 0.9995) with the slope of approaching unity (within 1.0509 and 1.1313). Conclusions: Data from the measurement of reference materials showed that the precision and accuracy of our method were satisfactory. This method is a powerful tool for in situ sulfur isotope measurement of sulfides and can be further applied to in situ carbon and oxygen isotope analyses. [ABSTRACT FROM AUTHOR]

The article presents a study of geochronological study of the Xuanlong Type Sedimentary Iron Deposits in North China Craton (NCC) by utilizing evidence from uranium-lead (U-Pb) dating ages of detrital zircons from the Chuanlinggou Formation, which is the hanging wall of iron ore bodies in the Jiangjiazhai and Pangjiapbu mines in China. It also states that banded iron formations (BIFs) represents the most important Precambrian mineralization in NCC.

Abstract The Huoqiu iron deposit is one of the few Superior-type BIF ore fields in China. To clarify the genesis of Huoqiu BIFs, we give out the geochemical and O-S isotopic data of the BIF samples and zircon Hf isotopic data of the wall-rock. The Huoqiu BIFs show dominant composition of SiO 2 + Fe 2 O 3 T, and varying contents of Al 2 O 3 , MgO, CaO, Rb, Sr, Zr and Hf, suggesting their formation mainly through chemical precipitation and incorporation of terrigenous felsic clastics. Shale-normalized rare earth element patterns of the BIF samples show the features characteristic of other Archean and Paleoproterozoic BIFs, with HREE enrichment relative to LREE, positive La and Eu anomalies, and superchondritic Y/Ho ratios comparable to modern seawater. Positive Eu anomalies are attributed to an imprint of high-temperature hydrothermal fluids. None to negative Ce anomaly shown by the BIFs imply the Huoqiu BIFs was precipitated from the mildly oxidized seawater in a nearshore setting. The quartz separated form Huoqiu BIF has δ18O V-SMOW values of 11.8‰–14.8‰, within the extent of the siliceous rocks deposited from high-temperature hydrothermal. The δ34S values of pyrite grains vary from −2.84 to +4.38‰, which is similar as the sulfur derived from mixtures between mantle fluids and sediments; The Δ33S values varied from −0.08‰ to +1.03‰, imply that the Huoqiu BIFs deposited distally from the volcanic center. The Hf model ages of these detrital zircons from the wall rock give three prominent peak at 3.1–3.0 Ga, 2.9–2.7 Ga and 2.5 Ga, respectively, it suggest that there were three episodes of the crust growth in the Huoqiu area. Integrated these data with the pioneering studies on the Huoqiu BIFs and wall rocks, we confirmed that the Huoqiu BIFs were sourced from the high-temperature hydrothermal related with the volcanics and precipitated from the mildly oxidized seawater in a nearshore setting and distally from the volcanics at 2.5 Ga. Highlights • Huoqiu BIFs sourced from hydrothermal fluids; • It precipitated from the mildly oxidized seawater; • It depoisted distally from the volcanics. [ABSTRACT FROM AUTHOR]

Background: Intestinal health plays a pivotal role in broiler chicken growth. Oregano aqueous extract (OAE) effectively exerts anti-inflammatory and antibacterial effects. However, the protective effects of OAE on intestinal health in broilers and the underlying mechanism remain unclear. This study aimed to investigate the potential effects of OAE on growth performance, the gut microbiota and intestinal health. A total of 840 1-d-old male and female broilers (Arbor Acres) were randomly allocated into 6 groups as follows: basal diet (Con), Con + antibiotics (Anti, colistin sulfate 7 g/kg, roxarsone 35 g/kg), Con + 400, 500, 600 and 700 mg/kg OAE (OAE400, OAE500, OAE600 and OAE700). Subsequently, fermentation in vitro together with oral administration trials were carried out to further assess the function of OAE on intestinal health of broilers. Results: Dietary 700 mg/kg OAE supplementation resulted in an increase (P < 0.05) in body weight and a decrease (P < 0.05) in feed conversion ratio when compared with the control during d 22 to 42 of the trial. OAE addition resulted in lower (P < 0.05) jejunal crypt depth and mRNA expression of IL-4 and IL-10 at d 42. In addition, dietary OAE addition increased the abundance of Firmicutes (P = 0.087) and Lactobacillus (P < 0.05) in the cecum, and increased (P < 0.05) the content of acetic acid and butyric acid. In the in vitro fermentation test, OAE significantly increased (P < 0.05) the abundance of Lactobacillus, decreased (P < 0.05) the abundance of unspecified_Enterobacteriaceae, and increased the content of acetic acid (P < 0.05). In the oral administration trial, higher (P < 0.05) IL-4 expression was found in broilers when oral inoculation with oregano fermentation microorganisms at d 42. And SIgA content in the ileum was significantly increased (P = 0.073) when giving OAE fermentation supernatant. Conclusions: Dietary OAE addition could maintain intestinal health and improve growth performance through enhancing intestinal mucosal immunity and barrier function mediated by gut microbiota changes. [ABSTRACT FROM AUTHOR]

Parentage identification technology is of great significance in the selective breeding of aquatic animals. Procambarus clarkii, at present, is an important freshwater economic species in China, and its genetic breeding is quite urgent for its aquaculture. In this study, 18 novel polymorphic microsatellite markers were developed for P. clarkii from 144 potential primers. The efficiency of 16 developed tetrameric markers with higher PIC values and two published tetrameric markers for parentage identification was evaluated in 15 maternal half-sib families. Ten highly polymorphic markers (mean HE = 0.720 and PIC = 0.669) were identified as being suitable for inclusion in the parentage marker suite. Simulation analysis indicated that the cumulative exclusion probabilities for the ten loci were 97.55% and 99.86%, respectively, when no parental information or only one parental information was available. In actual parentage assignments, 94.67% of offspring were correctly assigned to their mothers, which confirmed the application value of this marker suite in the parentage identification of P. clarkii. [ABSTRACT FROM AUTHOR]

The Huoqiu banded iron formation (BIF) iron deposits in the southeastern margin of the North China Craton (NCC) is hosted by middle-grade metamorphic Huoqiu complex. LA-ICP-MS U–Pb dating on zircons separated from the wallrock of the orebody constrains the depositional age ≤ 2.54 Ga and metamorphic age as ∼1.8 Ga. The dominant composition of SiO 2 + Fe 2 O 3 T of the BIF ores suggests their formation mainly through chemical precipitation. The widely varying contents of major elements such as Al 2 O 3 , MgO, CaO and trace elements such as Rb, Sr, Zr, Hf, Cr, Co, and Ni clearly indicate the incorporation of clastics, especially continental felsic clastics. When compared with Post-Archean Australian Shale (PAAS), the seawater-like signatures of REE distribution patterns, such as LREE depletion, positive La and Y anomalies, and superchondritic Y/Ho ratios, support the deposition in seawater. Meanwhile, the presence of strong positive Eu anomalies suggest the participation of hydrothermal fluids, which is according with the high depletion of 30 Si in Huoqiu BIFs. Magnetite and specularites within the BIFs contain heavy Fe isotopes resulting from the partial oxidation and precipitation of Fe 2+ to Fe 3+ in seawater. Integrated with geochronological and geochemical data from other Archean rocks in this area, we propose the formation of the BIFs occurred in a back-arc basin environment, and belonged to Superior-type. [ABSTRACT FROM AUTHOR]

The traditional method for sulfur isotope measurement using EA‐IRMS commonly requires sulfur content greater than 2 μmol. Such a large sample size limits its application to low‐S materials, the size mainly being due to ineffective utilization of sample gas, almost 99.7% of which is discarded with carrier gas through the split port of the continuous‐flow interface. A modified EA‐IRMS system with a gas chromatographic (GC) column and a custom‐built cryogenic concentration device is used in this study. We measured six reference materials to test the performance of this method. The results were consistent with those obtained through traditional EA‐IRMS. Precisions ranging from ±0.24‰ to ±0.76‰ (1σ) can be obtained with samples equivalent to ∼80 nmol sulfur, which were similar to results obtained from an alternative method using an absorption column. Our improved method is a powerful tool for sulfur isotope measurement in ultrasmall sulfide and sulfate samples, which can be further applied to carbon, nitrogen and oxygen isotope analyses of samples at about 100 nmol level. [ABSTRACT FROM AUTHOR]

Insulin-like androgenic gland factor (IAG) plays an important role in sex manipulation in decapods. Understanding the molecular regulation mechanism of IAG in Procambarus clarkii (PcIAG) is important for realizing its sex control. In this study, the promoter and gene structure of PcIAG, mRNA, and miRNA expression profiles after interfering with two siRNAs synthesized according to the two short repeats in the 5′ untranslated regions (5′UTR) of PcIAG were analyzed, and miRNAs of exosomes were investigated to explore the role of repeated sequences with tandem two short repeats located in the 5′UTR of PcIAG isolated from the androgenic gland (AG) in the regulation of IAG expression. The results showed that the repeated sequences of 5′UTR only occurred completely in the cDNA from AG, and the function of the two repeats was different in regulating the expression of PcIAG, in which the Wnt signaling pathway may be involved. Furthermore, we found that six miRNAs including miR-133, miR-193, miR-34, miR-1, miR-100, and let-7 might be involved in the regulation of the expression of PcIAG, wherein miR-133 might directly be related with the repeated sequences of 5′UTR. [ABSTRACT FROM AUTHOR]

In order to study the permeability evolution law of coal and rock under the action of mining dynamic load, the Darcy law steady-state measurement method was adopted to conduct the stress-strain permeability experiment of coal and rock using the triaxial compression permeability experiment device. The experimental results show that the increase of confining pressure prevents the development of tensile crack, resulting in the increase of stress peak and permeability decrease of the total stress-strain curve of coal and rock. With the same confining pressure, the variation trend of the coal rock strain-permeability curve and the stress-strain curve is basically the same, and there is a causal relationship, resulting in the hysteresis of the coal rock strain-permeability curve compared with the stress-strain curve.

Increasing evidence suggests that in vitro fertilization (IVF) may be associated with an increased risk of developing obesity and metabolic diseases later in life in the offspring. Notably, the addition of melatonin to culture medium may improve embryo development and prevent cardiovascular dysfunction in IVF adult mice. This study aimed to determine if melatonin supplementation in the culture medium can reverse impaired glucose metabolism in IVF mice offspring and the underlying mechanisms. Blastocysts used for transfer were generated by natural mating (control group) or IVF with or without melatonin (10−6 M) supplementation (mIVF and IVF group, respectively) in clinical‐grade culture media. Here, we first report that IVF decreased hepatic expression of Fbxl7, which was associated with impaired glucose metabolism in mice offspring. Melatonin addition reversed the phenotype by up‐regulating the expression of hepatic Fbxl7. In vitro experiments showed that Fbxl7 enhanced the insulin signaling pathway by degrading RhoA through ubiquitination and was up‐regulated by transcription factor Foxa2. Specific knockout of Fbxl7 in the liver of adult mice, through tail intravenous injection of recombinant adeno‐associated virus, impaired glucose tolerance, while overexpression of hepatic Fbxl7 significantly improved glucose tolerance in adult IVF mice. Thus, the data suggest that Fbxl7 plays an important role in maintaining glucose metabolism of mice, and melatonin supplementation in the culture medium may rescue the long‐term risk of metabolic diseases in IVF offspring. [ABSTRACT FROM AUTHOR]

[Display omitted] • Mafic microgranular enclaves were the product of mafic and felsic magma mixing. • Tieshan batholith originated from partial melting of enriched lithospheric mantle. • Skarn-porphyry Cu deposits prefer thicker crust whereas Fe skarns prefer normal one. Mineralogy, geochemistry, and geochronology of the mafic microgranular enclaves (MMEs) and their host diorite from the Tieshan batholith, Edong district, eastern China, were studied to constrain their petrogenesis and relationship with Fe-Cu skarn mineralization. The MMEs are diorite in composition and characterized by mixed apatite morphologies (elongated thin needles and stubby prismatic form) and bladed biotite, as well as clots of host diorite and megacrystic alkali-feldspar xenocrysts. The MMEs have similar zircon U-Pb age as their host diorite (~139 Ma). Most of the major and trace elements of the MMEs show a linear trend on Harker diagrams and plot along the low-silica trend of the host diorite. They are also characterized by enrichment of LREE and large ion lithophile elements (LILE), but are depleted in high field strength elements (HFSE). The MMEs display similar initial Sr isotopes and ε Nd (t), and slightly lower ε Hf (t) compared to that of host diorite. These results indicate that the Tieshan batholith formed via partial melting of an enriched lithospheric mantle, which is believed to be in equilibrium with a garnet-bearing residue (>50 km), followed by contamination of low crustal materials and fractional crystallization, whereas the MMEs appear to be the products of magma mixing. The presence of Cu- and Ni-bearing minerals in the MMEs probably indicates upper mantle or lower crust accumulation of sulfide which may have played a key role in controlling the formation of the parental magma associated with the skarn-porphyry Cu deposit (148 to 135 Ma) from the Middle-Lower Yangtze River metallogenic belt (MLYRMB). In contrast, the thinner, normal crust (~30 km) at ~ 130 Ma, believed to be in equilibrium with a plagioclase-rich residue, was either unsuitable for producing Cu-bearing accumulations or failed to transfer the deep accumulations to shallower levels to form Cu-bearing skarn systems, resulting in Fe-only mineralization. This may imply that skarn-porphyry Cu deposits prefer to form under > 50 km crustal settings, whereas Fe skarns may preferentially occur under normal (~30 km) crustal thickness. [ABSTRACT FROM AUTHOR]

The direct discharge of industrial emulsified wastewater critically threatens the environment and human health. Emulsified wastewater must be demulsified before discharge. Inspired by the applications of microchannels in oil-in-water (O/W) emulsion separation and patterned wettable surfaces to manipulate droplet formation, an arrow-shaped patterned microchannel (APM) was designed to enhance the droplet coalescence and demulsification of O/W emulsions with high oil content. The APM demulsification efficiency with 15 arrows of 5 patterns attained 70 % within 4.5 s when handling the O/W emulsion of 40 % (v / v) oil content. The experimental and simulation results revealed that the arrow-shaped patterns provided a series of inducing effects for dispersed droplets, enabling the coalesced droplets to form "oil-droplet-string flows". These positive inductions improved the efficiency of droplet coalescence and phase separation, thereby markedly enhancing the demulsification effect. By introducing the arrow-shaped wettable patterns, APM achieves the highest single demulsification efficiency of 70 % known at present for surfactant stabilized O/W emulsion with high oil content. This study provides a new technical approach for the treatment of industrial emulsified wastewater with high oil content. [Display omitted] • Arrow-shaped patterns enhance droplet coalescence efficiency and induce droplets to form "oil-droplet string flows". • Demulsification efficiency of O/W emulsion with 40 % (v / v) oil content attains 70 % in 4.5 s. • The demulsification efficiency of 5@15APM increases by 200 % compared to that of SHLM. • Numerical simulation explores the coalesce of droplets in microchannels of different wettabilities and heights. [ABSTRACT FROM AUTHOR]

The red swamp crayfish, Procambarus clarkii (Girard, 1852), is currently an economically important aquaculture animal. Its genetic basis has been scarcely reported, however, partly due to the absence of abundant molecular markers in the genome. In this study, Simple Sequence Repeat (SSR) loci were mined, based on genome survey sequencing via the next generation sequence of the red swamp crayfish. A total of 4897 SSR loci were identified, with the most abundant type being the di-nucleotide repeat motifs (75.2%), followed by tri- (20.4%), tetra- (3.8%), penta- (0.5%), and hexanucleotide (0.2%) repeats. In total, 1546 SSR markers were validated to be amplified, and 721 of these were identified as polymorphic SSR markers. Fifty polymorphic SSR markers were randomly selected for the identification of the genetic diversity of the 14 red swamp crayfish populations in China. The expected and observed heterozygosity and polymorphism information content were 0.39, 0.30, and 0.29, respectively, on average. The results indicated a medium genetic diversity among the 14 investigated populations. These probably cluster into three genetic populations. The current study provides abundant genetic markers and information on the 14 populations, which can be helpful for genetic diversity estimation and molecular breeding of the red swamp crayfish. Résumé: L'écrevisse rouge de Louisiane, Procambarus clarkii (Girard, 1852), est une espèce d'aquaculture importante économiquement. Sa base génétique est peu connue, due en partie à l'absence de marqueurs moléculaires abondants dans son génome. Dans cette étude, l'analyse des microsatellites (SSR) de l'écrevisse rouge a été effectuée, sur la base d'un séquençage du génome par séquençage nouvelle génération (NGS). Un total de 4897 loci SSR ont été identifiés, le type le plus abondant de motifs répétés étant les di-nucléotides (75,2%), suivi par les répétitions à tri- (20,4%), tétra- (3,8%), penta- (0,5%), et hexa-nucléotides (0,2%). Au total, 1546 marqueurs ont été validés pour être amplifiés, et 721 d'entre eux ont été identifiés comme marqueurs SSR polymorphiques. 50 marqueurs SSR polymorphiques ont été sélectionnés au hasard pour l'identification de la diversité génétique des 14 populations d'écrevisse rouge de Louisiane en Chine. L'hétérozygotie attendue et observée et l'indice de polymorphisme ont été, en moyenne, respectivement de 0,39, 0,30, et 0,29. Les résultats montrent une diversité génétique moyenne parmi les 14 populations étudiées, qui se regroupent probablement dans trois populations génétiques. Cette étude fournit des marqueurs génétiques abondants et des informations sur les 14 populations, qui peuvent être utiles pour l'estimation de la diversité génétique et la sélection moléculaire de l'écrevisse rouge de Louisiane. [ABSTRACT FROM AUTHOR]

Purpose: Tubulin beta eight class VIII (TUBB8) is essential for oogenesis, fertilization, and pre-implantation embryo development in human. Although TUBB8 mutations were recently discovered in meiosis-arrested oocytes of infertile females, there is no effective therapy for this gene mutation caused infertility. Our study aims to further reveal the infertility-causing gene mutations in the patient's family and to explore whether the infertility could be rescued by optimizing the conditions of embryo culture and finally achieve the purpose of making the patient pregnant. Methods: Whole-exome sequence analysis and Sanger sequencing were performed on patients' family members to screen and identify candidate mutant genes. Construction of plasmids, in vitro transcription, microinjection of disease-causing gene cRNA, and immunofluorescence staining were used to recapitulate the infertility phenotype observed in patients and to understand the pathogenic principles. Simultaneously, overexpression of mutant and wild-type cRNA of the candidate gene in mouse oocytes at either germinal vesicle (GV) or metaphase II (MII) stage was performed in the rescue experiment. Results: We first identified a novel heritable TUBB8 mutation (c.1041C>A: p.N347K) in the coding region which specifically affects the first mitosis and causes the developmental arrest of early embryos in a three-generation family. We further demonstrated that TUBB8 mutation could lead to abnormal spindle assemble. And moreover, additional expression of wild-type TUBB8 cRNA in the mouse oocytes in which the mutant TUBB8 were expressed can successfully rescue the developmental defects of resulting embryo and produce full-term offspring. Conclusions: Our study not only defines a novel mutation of TUBB8 causing the early cleavage arrest of embryos, but also provides an important basis for treating such female infertility in the future. [ABSTRACT FROM AUTHOR]

Carbohydrate sulfotransferases 11 (chst11) is one of the enzymes that synthesize chondroitin sulfate (CS), which has extensive immune functions in vitro and plays a critical role in mediating the infection of host by pathogenic microorganisms. However, whether it has immune functions in crayfish is still poorly understood. In our previous study of transcriptome, chst11 was differentially expressed in susceptible individuals and resistant individuals of Procambarus clarkii after white spot syndrome virus (WSSV) injection. Thus, in this study, the sequence of chst11 was obtained from P. clarkii for the first time and analyzed, and the expression pattern of chst11 was investigated. Besides, the purified recombinant protein of chst11 effect in protection in WSSV infection was explored. The full length of chst11 was 1536 bp with an 831-bp open reading frame (ORF), which encoding 276 amino acids residues with a calculated molecular mass of 33.1 kDa. The chst11 contains a Sulfotransfer_2 domain, one N -glycosylation site and three O-glycosylation sites. Phylogenetic analysis results showed that chst11 had the highest similarity to Penaeus vannamei (79.93%). The expression pattern of chst11 in different tissues indicated that chst11 was expressed highest in gut, gill and hypodermis, lowest in testicular duct, periesophageal nerve and hemocytes. The chst11 had different expression patterns in different tissues when the crayfish was challenged by WSSV, Aeromonas hydrophila and CpG ODN. Recombinant chst11 protein significantly reduced the amount of WSSV copy number in hepatopancreas at 6 h and 12 h post injection compared to the control group injected with bovine serum albumin (BSA). It was found that chst11 protein enhanced the expression of peroxinectin , proPO in hepatopancreas and midgut and the C-type lectin (ctl) in hemocytes and hepatopancreas. Intramuscularly injection of juvenile crayfish with chst11 protein decreased 60% mortality compared to the control group with BSA. This study is the first report on the antiviral function of chst11 in the immune system of crustacean. • The chst11 gene was isolated from Procambarus clarkii. • Chst1 1 had different expression patterns in different tissues. • The chst11 gene had the antiviral function. • Procambarus clarkii chst11 enhanced resistance to WSSV. • Recombinant chst11 enhanced the mRNA expression of peroxinectin , proPO and ctl. [ABSTRACT FROM AUTHOR]

Copper and iron skarn deposits are economically important types of skarn deposits throughout the world, especially in China, but the differences between Cu and Fe skarn deposits are poorly constrained. The Edong ore district in southeastern Hubei Province, Middle–Lower Yangtze River metallogenic belt, China, contains numerous Fe and Cu–Fe skarn deposits. In this contribution, variations in skarn mineralogy, mineralization-related intrusions and sulfur isotope values between these Cu–Fe and Fe skarn deposits are discussed. The garnets and pyroxenes of the Cu–Fe and Fe skarn deposits in the Edong ore district share similar compositions, i.e., dominantly andradite (Ad 29–100 Gr 0–68 ) and diopside (Di 54–100 Hd 0–38 ), respectively. This feature indicates that the mineral compositions of skarn silicate mineral assemblages were not the critical controlling factors for variations between the Cu–Fe and Fe skarn deposits. Intrusions associated with skarn Fe deposits in the Edong ore district differ from those Cu–Fe skarn deposits in petrology, geochemistry and Sr–Nd isotope. Intrusions associated with Fe deposits have large variations in their (La/Yb) N ratios (3.84–24.6) and Eu anomalies (δEu = 0.32–1.65), and have relatively low Sr/Y ratios (4.2–44.0) and high Yb contents (1.20–11.8 ppm), as well as radiogenic Sr–Nd isotopes (εNd(t) = − 12.5 to − 9.2) and ( 87 Sr/ 86 Sr) i = 0.7067 to 0.7086. In contrast, intrusions associated with Cu–Fe deposits are characterized by relatively high Sr/Y (35.0–81.3) and (La/Yb) N (15.0–31.6) ratios, low Yb contents (1.00–1.62 ppm) without obvious Eu anomalies (δEu = 0.67–0.97), as well as ( 87 Sr/ 86 Sr) i = 0.7055 to 0.7068 and εNd(t) = − 7.9 to − 3.4. Geochemical evidence indicates a greater contribution from the crust in intrusions associated with Fe skarn deposits than in intrusions associated with Cu–Fe skarn deposits. In the Edong ore district, the sulfides and sulfates in the Cu–Fe skarn deposits have sulfur isotope signatures that differ from those of Fe skarn deposits. The Cu–Fe skarn deposits have a narrow range of δ 34 S values from − 6.2‰ to + 8.7‰ in sulfides, and + 13.2‰ to + 15.2‰ in anhydrite, while the Fe skarn deposits have a wide range of δ 34 S values from + 10.3‰ to + 20.0‰ in pyrite and + 18.9‰ to + 30.8‰ in anhydrite. Sulfur isotope data for anhydrite and sedimentary country rocks suggest that the formation of skarns in the Edong district involved the interaction between magmatic fluids and variable amounts of evaporites in host rocks. [ABSTRACT FROM AUTHOR]

In hydrothermal high-temperature abnormal mines, the composite heat-insulation zone structure, formed through a combination of guniting and grouting, serves to mitigate heat dissipation from the surrounding rock into the airflow. To comprehensively understand the thermal insulation performance of the composite heat-insulation zone structure, this study employs numerical simulation to analyze the following aspects: the variation in the temperature field within the surrounding rock of the roadway without insulation, the influence of structural parameters of the composite heat-insulation zone on temperature distribution in the surrounding rock of the roadway, and the thermal insulation effectiveness of the composite heat-insulation zone with varying structures. The findings indicate that the temperature distribution within the surrounding rock of the roadway lacking a heat-insulation zone is relatively uniform. However, as ventilation time extends, the heat regulation zone within the surrounding rock gradually extends deeper, ultimately forming an elliptical cooling area. The composite heat-insulation zone structure effectively mitigates heat transfer from deeper surrounding rock to the roadway wall, consequently altering the scope of the roadway's heat regulation zone. Enhancing the thermal insulation performance of the composite heat-insulation zone structure can be achieved by increasing the thickness of the thermal insulation layer, adjusting grouting rate and depth, and reducing the thermal conductivity of insulation materials. The thermal insulation effectiveness of the thermal insulation layer surpasses that of the grouting layer, with its performance primarily influenced by the thermal conductivity of the materials used. Simulation results demonstrate that the composite heat-insulation zone structure reduces the maximum heat flux on the roadway wall from 47.4 to 37.7 W/m 2 , resulting in a 20% reduction in heat transfer from deeper surrounding rock. These findings offer valuable insights for implementing thermal insulation techniques in hydrothermal high-temperature anomaly mines., (© 2024. The Author(s).)

Growth is a crucial economic trait of all aquaculture species. It is important to explore the molecular regulation on growth, which could help improve the growth rate of species. Mining the growth-related genes is the foundation for revealing its molecular regulation on growth. Presently, the molecular regulation of growth in Procambarus clarkii is not clear, and the study on exploring growth-related genes is limited. In this study, RNA-Seq was used to compare gene expression profiles of the individuals with different growth rates involved in four groups including Big Male (BM), Big Female (BF), Small male (SM), and Small Female (SF) from one P. clarkii family, and the analyses were performed in combination with sex. Meanwhile, whole-genome resequencing data was used to get growth-specific SNP (Single Nucleotide Polymorphism)/InDel (Insertion/Deletion) sites information. Totally, we identified 16,127 genes, of which 9065 were successfully annotated in the GO database. Among these, 1328 DEGs were identified in BM vs. SM, with 357 up-regulated and 971 down-regulated. Additionally, 3507 DEGs were identified in BF vs. SF, with 241 up-regulated and 3266 down-regulated. 96 DEGs were up-regulated and 820 DEGs were down-regulated in Growth-related Group. The expression levels of nine DEGs were validated by RT-qPCR to verify the analysis results of sequencing. 684,040 growth-related SNPs and 182,050 growth-related InDels were obtained after screened. These findings provide candidate growth-related genes and growth-specific SNP/InDel sites for regulation of growth traits in P. clarkii, and new insight into the molecular regulation of P. clarkii growth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The insulin-like androgenic gland hormone gene (IAG), primarily expressed in the androgenic gland (AG), plays a crucial role in controlling male sex differentiation and maintaining male secondary sexual characteristics in decapods. In this study, we investigated the mRNA and microRNA expression profiles of male Procambarus clarkii to understand the transcriptomic regulatory mechanism of IAG after the injection of an efficient siRNA (GsiRNA) designed based on IAG. The results revealed that several differentially expressed genes were enriched in reproduction-related pathways, such as the wnt signaling pathway, MAPK signaling pathway, and GnRH signaling pathway. In the testis (Te), the injection of GsiRNA led to the up-regulation of many ovary-related genes and down-regulation of testis-related genes. Moreover, the brain (Br) and abdominal nerve cord (AN) appeared to be involved in the regulation of IAG, with numerous differentially expressed genes found in Br and AN. Notably, the expression of five neuropeptide genes, Crustacean hyperglycemic hormone, pigment-dispersing hormone, red pigment concentrating hormone precursor, corazonin, and gonadotropin-releasing hormone II receptor isoform X1 in Br/AN, was significantly changed. Additionally, three ovary-related miRNAs (miR-263a, miR-263b, miR-133) highly expressed in Te/AG showed significant up-regulation after GsiRNA injection. Furthermore, the long-term interference of GsiRNA was found to inhibit the development of male external sexual characteristics during the juvenile stage and delay it during the adult stage. This research provides valuable insights into the molecular regulatory mechanism and function of IAG in P. clarkii., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Somatic cell nuclear transfer (SCNT) can reprogram differentiated somatic cells into totipotency. Although pre-implantation development of SCNT embryos has greatly improved, most SCNT blastocysts are still arrested at the peri-implantation stage, and the underlying mechanism remains elusive. Here, we develop a 3D in vitro culture system for SCNT peri-implantation embryos and discover that persistent Wnt signals block the naïve-to-primed pluripotency transition of epiblasts with aberrant H3K27me3 occupancy, which in turn leads to defects in epiblast transformation events and subsequent implantation failure. Strikingly, manipulating Wnt signals can attenuate the pluripotency transition and H3K27me3 deposition defects in epiblasts and achieve up to a 9-fold increase in cloning efficiency. Finally, single-cell RNA-seq analysis reveals that Wnt inhibition markedly enhances the lineage developmental trajectories of SCNT blastocysts during peri-implantation development. Overall, these findings reveal diminished potentials of SCNT blastocysts for lineage specification and validate a critical peri-implantation barrier for SCNT embryos., (© The Author(s) 2023. Published by Oxford University Press on behalf of China Science Publishing & Media Ltd.)

As an important energy base in central China, the Pingdingshan coalfield has abundant coal and geothermal resources. The cooperative exploration of coal and geothermal resources is significant for the comprehensive utilization of energy resources. This work collected coal-bearing samples from the Pingdingshan coalfield to investigate the tectono-thermal evolution of a high geothermal coalfield, especially the present geothermal field and hydrocarbon generation model. The geochemical results show that the Shanxi and Taiyuan source rocks have average R o values of 0.88 and 0.97%, respectively, with an average Rock-Eval T max value of 442 °C. Hydrocarbon generation of source rocks started at ∼205 Ma, with the highest rates at ∼170 Ma, reaching the maximum transformation ratio of 40-50% in the middle of the Early Cretaceous. The age and length of apatite fission tracks (AFTs) indicate that coal-bearing strata underwent significant post-depositional annealing after the Late Permian and suggest an abnormal thermal event that occurred in the Late Mesozoic. Meso-Cenozoic thermal event was mainly caused by the plutonic metamorphism of the Early Jurassic and magmatic thermal metamorphism of the Early Cretaceous, achieving a maximum paleotemperature of ∼140 °C. The magmatic thermal event resulted from the intensive post-orogenic extension of the Qinling-Dabie Orogenic Belt caused by the tectonic transition of the North and South China Plates. The present-day high geotemperature of Pingdingshan Coalfield is dominated by the horst structure caused by the regional extension of the basin-mountain system. The Cambrian limestone with a high thermal conductivity underlying coal measure collects deep heat, forming a heat accumulation center of this horst structure with a heat flow of 74 mW/m 2 and a maximum temperature of ∼50 °C nowadays., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro., (©The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.)

Background: Successful cause-related marketing (CRM) campaigns can help companies stand out from their competitors; however, CRM may not have pleasant outcomes, even if it receives substantial investment., Objective: This research aimed to investigate how gamified CRM projects influence consumers' favorability., Methods: We introduced 3 different CRM projects in 3 different studies. Every project had 2 versions according to the level of gamification, and participants were randomly assigned into these 2 groups. Additionally, we used a 2 (gamification: lower, higher) 2 (rules presentation: without visual cues, with visual cues) between-subjects design to test the moderation role of rules presentation in gamified CRM projects., Results: In Study 1, we identified that the highly gamified CRM program induces more enjoyment (F 1,139 =21.11, P<.001) and higher favorability (F 1,139 =14.57, P<.001). Moreover, we found that enjoyment played a mediation role between gamification and favorability (P<.001) in Study 2. In addition, the results of Study 3 indicated rules presentation in a gamified CRM program can moderate the indirect effect of gamification on favorability via enjoyment (index of the moderated mediation: 95% CI -1.12 to -0.10; for rules presentation with visual cues: 95% CI 0.69 to 1.40; for rules presentation without visual cues: 95% CI 0.08 to 0.83)., Conclusions: Overall, this research contributes to the CRM literature and suggests gamification is an effective way of managing CRM campaigns., (©Yanhe Li, Yanchen Li, Xiu Zhou, Kunshu Ma. Originally published in JMIR Serious Games (https://games.jmir.org), 10.01.2023.)

As a new form of online reviews, Q&A reviews have been recently used by many e-commerce platforms to compensate for the weaknesses and problems related to trust and helpfulness found in traditional online reviews. This research documents what motivates people to share products or purchasing knowledge with others through Q&A reviews and why e-commerce platforms should place an emphasis on Q&A reviews. Importantly, our results provide evidence that, when receiving feedback (i.e., comments and likes), people are more likely willing to share knowledge with others and will have a higher level of loyalty. We believe that this study contributes to knowledge sharing and the e-commerce literature, and also has practical implications., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Li, Li, Ma and Zhou.)

N 6 -methyladenosine (m 6 A) and its regulatory components play critical roles in various developmental processes in mammals. However, the landscape and function of m 6 A in early embryos remain unclear owing to limited materials. Here we developed a method of ultralow-input m 6 A RNA immunoprecipitation followed by sequencing to reveal the transcriptome-wide m 6 A landscape in mouse oocytes and early embryos and found unique enrichment and dynamics of m 6 A RNA modifications on maternal and zygotic RNAs, including the transcripts of transposable elements MTA and MERVL. Notably, we found that the maternal protein KIAA1429, a component of the m 6 A methyltransferase complex, was essential for m 6 A deposition on maternal mRNAs that undergo decay after zygotic genome activation and MTA transcripts to maintain their stability in oocytes. Interestingly, m 6 A methyltransferases, especially METTL3, deposited m 6 A on mRNAs transcribed during zygotic genome activation and ensured their decay after the two-cell stage, including Zscan4 and MERVL. Together, our findings uncover the essential functions of m 6 A in specific contexts during the maternal-to-zygotic transition, namely ensuring the stability of mRNAs in oocytes and the decay of two-cell-specific transcripts after fertilization., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

N 6 -methyladenosine (m 6 A) is the most abundant internal modification on mammalian messenger RNA. It is installed by a writer complex and can be reversed by erasers such as the fat mass and obesity-associated protein FTO. Despite extensive research, the primary physiological substrates of FTO in mammalian tissues and development remain elusive. Here, we show that FTO mediates m 6 A demethylation of long-interspersed element-1 (LINE1) RNA in mouse embryonic stem cells (mESCs), regulating LINE1 RNA abundance and the local chromatin state, which in turn modulates the transcription of LINE1-containing genes. FTO-mediated LINE1 RNA m 6 A demethylation also plays regulatory roles in shaping chromatin state and gene expression during mouse oocyte and embryonic development. Our results suggest broad effects of LINE1 RNA m 6 A demethylation by FTO in mammals.

Six novel verrucosidin derivatives, namely, poloncosidins A-F ( 1 - 6 ), together with one known analogue ( 7 ), were isolated and identified from the deep-sea-derived fungus Penicillium polonicum CS-252, which was obtained from cold-seep sediments collected in the South China Sea at a depth of 1183 m. Their structures were mainly established on the basis of a detailed interpretation of NMR spectroscopic and mass spectrometric data. The relative and absolute configurations of compounds 1 - 6 were determined by ECD calculations and a DP4+ probability analysis. Compounds 1 - 5 represent the first examples of verrucosidins with a 2,5-dihydrofuran ring which is uncommon among the known analogues. These compounds exhibited inhibitory activities against several human and aquatic pathogens with MIC values ranging from 4 to 32 μg/mL.

Parental DNA methylation and histone modifications undergo distinct global reprogramming in mammalian pre-implantation embryos, but the landscape of epigenetic crosstalk and its effects on embryogenesis are largely unknown. Here we comprehensively analyse the association between DNA methylation and H3K9me3 reprogramming in mouse pre-implantation embryos and reveal that CpG-rich genomic loci with high H3K9me3 signal and DNA methylation level (CHM) are hotspots of DNA methylation maintenance during pre-implantation embryogenesis. We further profile the allele-specific epigenetic map with unprecedented resolution in gynogenetic and androgenetic embryos, respectively, and identify 1,279 allele-specific CHMs, including 19 known imprinting control regions (ICRs). Our study suggests that 22 ICR-like regions (ICRLRs) may regulate allele-specific transcription similarly to known ICRs, and five of them are confirmed to be important for mouse embryo development. Taken together, our study reveals the widespread existence of allele-specific CHMs and largely extends the scope of allele-specific regulation in mammalian pre-implantation embryos., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Many economic crustacean species have sex dimorphisms during their growth. Exploring the sex determination system and developing sex-specific molecular marker(s) are very helpful for carrying out sex control breeding, and next-generation sequencing has been used as an efficient way to explore them in recent years. In this study, first, the genetic sex determination system of P. clarkii was explored as an XX/XY system by analyzing the 2b-RAD sequencing data. Furthermore, DNA samples of male and female individuals from a P. clarkii family were pooled separately for whole-genome resequencing. Based on the data of whole-genome resequencing, the 9,163 male- and female-specific bias sites with higher feasibility were obtained based on the assumption of the XX/XY sex determination system, and four sites were selected to design the sex-specific marker primers. One efficient sex-specific marker was identified with a sex discrimination rate of 99.49% (195/196) when applied to five different geographical groups with 196 individuals. The results of this study would provide a foundation for the realization of P. clarkii sex control and could provide some reference for investigating the sex determination system and sex molecular marker(s) of other crustacean species based on next-generation sequencing data., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Shen, Wang, Wang and Li.)

The histone variant H3.3 is an important maternal factor in fertilization of oocytes and reprogramming of somatic cell nuclear transfer (SCNT) embryos. As a crucial replacement histone, maternal H3.3 is involved in chromatin remodeling and zygote genome activation. Litte is, however, known about the replacement of H3.3 in the bovine SCNT embryos. In this study, the maternal H3.3 in mature ooplasm was labeled with HA tag and the donor cells H3.3 was labeled with Flag tag, in order to observe the replacement of H3.3 in the bovine SCNT embryos. Meanwhile, maternal H3.3 knockdown was performed by microinjecting two different interfering fragments before nucleus transfer. It was showed that the dynamic replacement between maternal- and donor nucleus-derived H3.3 was detected after SCNT. And it could be observed that the blastocyst development rate of the cloned embryos decreased from 22.3% to 8.2-10.3% (P < 0.05), the expression of Pou5f1 and Sox2 was down-regulated and the level of H3K9me3 was increased in the interfered embryos. In summary, H3.3 replacement impacted on the process of reprogramming, including embryonic development potential, activation of pluripotency genes and epigenetic modification in bovine SCNT embryos., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Poor oocyte quality is associated with early embryo developmental arrest and infertility. Maternal gene plays crucial roles in the regulation of oocyte maturation, and its mutation is a common cause of female infertility. However, how to improve oocyte quality and develop effective therapy for maternal gene mutation remains elusive. Here, we use Zar1 as an example to assess the feasibility of genome transfer to cure maternal gene mutation-caused female infertility. We first discover that cytoplasmic deficiency primarily leads to Zar1-null embryo developmental arrest by disturbing maternal transcript degradation and minor zygotic genome activation (ZGA) during the maternal-zygotic transition. We next perform genome transfer at the oocyte (spindle transfer or polar body transfer) and zygote (early pronuclear transfer or late pronuclear transfer) stages to validate the feasibility of preventing Zar1 mutation-caused infertility. We finally demonstrate that genome transfer either at the oocyte or at the early pronuclear stage can support normal preimplantation embryo development and produce live offspring. Moreover, those pups grow to adulthood and show normal fertility. Therefore, our findings provide an effective basis of therapies for the treatment of female infertility caused by maternal gene mutation., (Copyright © 2020 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.)

The insulin-like androgenic gland (IAG) gene is well known in male crustacean, and it is a key regulator in male sexual differentiation and maintaining the male sexual characteristic. The neo-female can be produced by silencing the MrIAG (Macrobrachium rosenbergii Insulin-like Androgenic Gland) in male Macrobrachium rosenbergii. This is the first time to use siRNA approach to silenced MrIAG in male M. rosenbergii. In the current study, the optimal injection dosage to achieve sex reversal is 0.5 μg/g body weight. After MrIAG silencing, the expression level of Dmrt11e, Dmrt99b, MRPINK, Mrr, Sxl1, and Sxl2 decreased significantly. As their long-term silencing effect of MrIAG, the dsRNA and siRNA approaches produce three and two individual neo-females, respectively. The neo-female has a wider brood chamber, ovipositing setae, and ovigerous setae, which is resembled normal female. After a long-term silencing with siRNA, most of the germ cells were arrested in spermatocytes stage, but the spermatocytes in control can further developed into spermatozoon. The seminiferous tubules are loosely arranged and the spermatocytes are more than spermatozoon in the 0.5 μg/g body weight treatment dose. This current study suggests a new path to obtain neo-females through siRNA silencing.

The oocyte cytoplasm can reprogram the somatic cell nucleus into a totipotent state, but with low efficiency. The spatiotemporal chromatin organization of somatic cell nuclear transfer (SCNT) embryos remains elusive. Here, we examine higher order chromatin structures of mouse SCNT embryos using a low-input Hi-C method. We find that donor cell chromatin transforms to the metaphase state rapidly after SCNT along with the dissolution of typical 3D chromatin structure. Intriguingly, the genome undergoes a mitotic metaphase-like to meiosis metaphase II-like transition following activation. Subsequently, weak chromatin compartments and topologically associating domains (TADs) emerge following metaphase exit. TADs are further removed until the 2-cell stage before being progressively reestablished. Obvious defects including stronger TAD boundaries, aberrant super-enhancer and promoter interactions are found in SCNT embryos. These defects are partially caused by inherited H3K9me3, and can be rescued by Kdm4d overexpression. These observations provide insight into chromatin architecture reorganization during SCNT embryo development.

DNA methylation and histone modifications critically regulate the expression of many genes and repeat regions during spermatogenesis. However, the molecular details of these processes in male germ cells remain to be addressed. Here, using isolated murine sperm cells, ultra-low-input native ChIP-Seq (ULI-NChIP-Seq), and whole genome bisulfite sequencing (WGBS), we investigated genome-wide DNA methylation patterns and histone 3 Lys-9 trimethylation (H3K9me3) modifications during mouse spermatogenesis. We found that DNA methylation and H3K9me3 have distinct sequence preferences and dynamics in promoters and repeat elements during spermatogenesis. H3K9me3 modifications in histones at gene promoters were highly enriched in round spermatids. H3K9me3 modification on long terminal repeats (LTRs) and long interspersed nuclear elements (LINEs) was involved in silencing active transcription from these regions in conjunction with reestablishment of DNA methylation. Furthermore, H3K9me3 remodeling on the X chromosome was involved in meiotic sex chromosome inactivation and in partial transcriptional reactivation of sex chromosomes in spermatids. Our findings also revealed the DNA methylation patterns and H3K9me3 modification profiles of paternal and maternal germline imprinting control regions (gICRs) during spermatogenesis. Taken together, our results provide a genome-wide map of H3K9me3 modifications during mouse spermatogenesis that may be helpful for understanding male reproductive disorders., (© 2019 Liu et al.)

Red swamp crayfish, Procambarus clarkii, presently is an important aquatic commercial species in China. The crayfish is a hot area of research focus, and its genetic improvement is quite urgent for the crayfish aquaculture in China. However, the knowledge of its genomic landscape is limited. In this study, a survey of P. clarkii genome was investigated based on Illumina's Solexa sequencing platform. Meanwhile, its genome size was estimated using flow cytometry. Interestingly, the genome size estimated is about 8.50 Gb by flow cytometry and 1.86 Gb with genome survey sequencing. Based on the assembled genome sequences, total of 136,962 genes and 152,268 exons were predicted, and the predicted genes ranged from 150 to 12,807 bp in length. The survey sequences could help accelerate the progress of gene discovery involved in genetic diversity and evolutionary analysis, even though it could not successfully applied for estimation of P. clarkii genome size.

Objective: To analyze the feasibility and effectiveness of fractionated Gamma Knife surgery (FGKS) for giant pituitary adenomas., Methods: From June 2005 to May 2016, 14 patients with giant pituitary adenomas were treated with FGKS, and 10 patients (71%) completed follow-up evaluation. All patients had undergone surgical resection at least once prior to FGKS. The median-volume of the adenomas was 17.6cm 3 (range 4.9-61cm 3 )., Results: The median follow-up period was 31.5 months (range 6-58 months). The size of the tumors decreased in 6 patients and remained stable in 4 patients. The visual acuity improved in 1 patient. None of the patients suffered from vision deterioration caused by FGKS., Conclusion: FGKS is an effective treatment modality for giant pituitary adenomas in selected patients., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Crayfish exoskeleton (CE) samples are generally less invasive and easy to be collected. However, it is difficult to extract DNA from them. This study was intended to investigate CE as a DNA source and design an easy and efficient DNA extraction protocol for polymerase chain reactions. Specific primer pair (PPO-F, PPO-R) was used to amplify extracted DNA from CE, and compared to crayfish tail muscle DNA sample. Moreover, seven microsatellites markers were used to amplify the CE DNA samples set. Since the extracted DNA from CE is suitable for gene amplification, the results present usefulness of CE as an easy and convenient DNA source for PCR-based population genetic research.

Accumulating evidence has indicated that epidermal growth factor receptor (EGFR) signaling pathway plays a potentially important role in mediating radiation resistance in human gliomas. In this study, we aimed to evaluate the effect of combination therapy with Gamma Knife radiosurgery (GKR) and antisense EGFR for malignant glioma in vitro and orthotopic xenografts. GKR and down-regulation of EGFR expression by antisense EGFR resulted in significant suppression of cell proliferation and induction of cell apoptosis in vitro. Moreover, the growth of the C6 glioma in orthotopic xenografts was significantly inhibited by this combination treatment. Thus, our results indicate that down-regulation of EGFR expression enhanced the radiosensitivity of glioma and GKR in combination with antisense EGFR is a potential strategy for glioma therapy.

Objective: To construct the small interfering RNA (siRNA) expression constructs targeting epidermal growth factor receptor(EGFR) and express them in TJ905 human malignant cells., Methods: Two target sequences from Receptor L domain and catalytic domain were selected to create two expression constructs using psiRNA-NeoG2. Furthermore, the siRNA constructs were transfected into TJ905 cells as mediated by Lipofectamin. Meanwhile, an antisense EGFR construct p-anti-hEGFR was set as control. Immunofluorescence and Western blot were performed to detect EGFR expression., Results: With the successful construction of the two siRNA expression plasmids and the stable transfection to TJ905 cells, the expression of EGFR was down-regulated to 90% and 92% respectively, but to 82% in the anti-sense EGFR group., Conclusion: The siRNA expression constructs targeting EGFR could specifically inhibit EGFR expression, and should be a new strategy in glioma gene therapy targeting EGFR.