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Abstract Background Improving glioma treatment effectiveness requires a thorough understanding of gliomagenesis. Emerging evidences have proved that zinc and ring finger 2 (ZNRF2) contributes to development of various human malignancies. Nevertheless, a comprehensive study of ZNRF2’s role in glioma is absent currently. Methods Utilizing open resources from Chinese Glioma Genome Atlas (CGGA), Gene Expression Omnibus (GEO), The Cancer Genome Atlas (TCGA), Connectivity Map (cMap) and other bioinformatic tools, we systematically examined the expression, clinical significance, prognostic value, regulated biological processes, immune infiltration, and potential inhibitors of ZNRF2 in gliomas. Functional experiments were also performed to validate its oncogenic roles in glioblastoma (GBM) cells. Results Our findings revealed that ZNRF2 expression was elevated in gliomas compared to normal brains, and its higher levels were correlated with increased grades and worse patient prognosis. The immune analysis suggested that immunotherapies targeting immune checkpoint genes could be beneficial for glioma patients with elevated ZNRF2 expression. Endogenous ZNRF2 knockdown impaired GBM cell proliferation, G2/M cell cycle progression and glycolysis, which was revealed by reduced ATP, pyruvic acid, lactic acid levels and less glucose uptake. Finally, we identified methylprednisolone (MP) as a potential ZNRF2 inhibitor and validated its anti-glioma effects in vitro. MP also enhanced the sensitization of GBM cells to temozolomide (TMZ), the primary chemotherapeutic agent for GBM in clinic. Conclusions Taken together, our study demonstrated ZNRF2 as an essential tumor-promoting factor by favoring GBM cell proliferation and glycolysis. Our findings suggested that ZNRF2 might serve as a novel independent prognostic biomarker and promising therapeutic target for glioma patients.

Hangzhou was the political and economic center of the Southern Song Dynasty (1127–1279 AD) and also the southern end of the Beijing-Hangzhou Grand Canal during the Ming and Qing Dynasties (1368–1644 AD). This historical position allowed the city’s economy to develop rapidly and influenced the form of its polychrome paintings with the imperial official style of the north China. However, due to the high temperature and rainy natural preservation conditions, southern polychrome paintings have always been a weak link in Chinese architectural polychrome painting craftsmanship. This study focuses on two well-preserved official-style architectural polychrome paintings in the grand halls from the late Qing period in Hangzhou. Through multi-techniques such as optical microscopy (OM), scanning electron microprobe with energy dispersive X-ray spectroscopy analysis (SEM-EDX), micro-Raman spectroscopy, micro-Fourier Transform Infrared spectroscopy (μ-FTIR), and pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), it was found that there is a significant difference from the reported common non-ground architectural paintings in the south, typically having four-layer structures with a white base and ground plaster layer in preparation for painting. The appearance of pigments such as artificial ultramarine (Na6Al4Si6S4O20) and emerald green (Cu(C2H3O2)2·3Cu(AsO2)2) indicates that the paintings were made at least after the 1830s, and the use of malachite green dye and copper phthalocyanine blue (PB 15:X) suggests that unrecorded restorations were also performed after the 20th century. All samples are coated with a layer of alkyd resin, which may have been added during the repairs in the latter half of the 20th century, leading to the black discoloration of the present paintings, especially in areas where emerald green was used. This study provides an important case for the study of the official style of polychrome painting craftsmanship in the southern region of China and also offers important references for the future protection and restoration of traditional architectural polychrome painting.

The contribution of plant hormones and energy-rich compounds and their metabolites (ECMs) in alleviating aluminum (Al) toxicity by elevated pH remains to be clarified. For the first time, a targeted metabolome was applied to identify Al-pH-interaction-responsive hormones and ECMs in Citrus sinensis leaves. More Al-toxicity-responsive hormones and ECMs were identified at pH 4.0 [4 (10) upregulated and 7 (17) downregulated hormones (ECMs)] than those at pH 3.0 [1 (9) upregulated and 4 (14) downregulated hormones (ECMs)], suggesting that the elevated pH improved the adaptation of hormones and ECMs to Al toxicity in leaves. The roles of hormones and ECMs in reducing leaf Al toxicity mediated by elevated pH might include the following aspects: (a) improved leaf growth by upregulating the levels of jasmonoyl-L-isoleucine (JA-ILE), 6-benzyladenosine (BAPR), N6-isopentenyladenosine (IPR), cis-zeatin-O-glucoside riboside (cZROG), and auxins (AUXs), preventing Al toxicity-induced reduction of gibberellin (GA) biosynthesis, and avoiding jasmonic acid (JA)-mediated defense; (b) enhanced biosynthesis and accumulation of tryptophan (TRP), as well as the resulting increase in biosynthesis of auxin, melatonin and secondary metabolites (SMs); (c) improved ability to maintain the homeostasis of ATP and other phosphorus (P)-containing ECMs; and (d) enhanced internal detoxification of Al due to increased organic acid (OA) and SM accumulation and elevated ability to detoxify reactive oxygen species (ROS) due to enhanced SM accumulation. To conclude, the current results corroborate the hypotheses that elevated pH reduces Al toxicity by upregulating the ability to maintain the homeostasis of ATP and other P-containing ECMs in leaves under Al toxicity and (b) hormones participate in the elevated pH-mediated alleviation of Al toxicity by positively regulating growth, the ability to detoxify ROS, and the internal detoxification of Al in leaves under Al toxicity. Our findings provide novel insights into the roles of hormones and ECMs in mitigating Al toxicity mediated by the elevated pH.

Abstract To investigate the boiling characteristics of flow outside the R410A tube under swaying conditions, this article conducts numerical simulation and experimental research on the flow boiling heat transfer of R410A outside a horizontal tube. The results show that when the swing frequency increased from 0.2 to 2 Hz, the sway amplitude is 0.03 m, the heat flux on the inner wall of the runner remains unchanged, and the mass flow rate increases from 85 to 170 kg/(m2·s), which makes the heat transfer coefficient of the working fluid in the annular area increases significantly. Keeping the inlet mass flow rate unchanged, the heat flux on the inner wall of the flow channel increases from 25 to 35 kW/m2, the heat transfer coefficient of the working fluid in the annular area has also improved, but under high heat flux conditions, the working fluid is evaporated and dried, its heat transfer coefficient increases less than in low heat flux conditions. When the sway amplitude increases from 0.02 to 0.07 m, the sway frequency is 0.2 Hz and 2 Hz respectively, and the heat transfer coefficient of the working fluid shows a downward trend as a whole. The studies provide a reference for heat exchanger design suitable for offshore swaying conditions.

Some citrus orchards in China often experience nitrogen (N) deficiency. For the first time, targeted metabolomics was used to examine N-deficient effects on hormones in sweet orange (Citrus sinensis (L.) Osbeck cv. Xuegan) leaves and roots. The purpose was to validate the hypothesis that hormones play a role in N deficiency tolerance by regulating root/shoot dry weight ratio (R/S), root system architecture (RSA), and leaf and root senescence. N deficiency-induced decreases in gibberellins and indole-3-acetic acid (IAA) levels and increases in cis(+)-12-oxophytodienoic acid (OPDA) levels, ethylene production, and salicylic acid (SA) biosynthesis might contribute to reduced growth and accelerated senescence in leaves. The increased ethylene formation in N-deficient leaves might be caused by increased 1-aminocyclopropanecarboxylic acid and OPDA and decreased abscisic acid (ABA). N deficiency increased R/S, altered RSA, and delayed root senescence by lowering cytokinins, jasmonic acid, OPDA, and ABA levels and ethylene and SA biosynthesis, increasing 5-deoxystrigol levels, and maintaining IAA and gibberellin homeostasis. The unchanged IAA concentration in N-deficient roots involved increased leaf-to-root IAA transport. The different responses of leaf and root hormones to N deficiency might be involved in the regulation of R/S, RSA, and leaf and root senescence, thus improving N use efficiency, N remobilization efficiency, and the ability to acquire N, and hence conferring N deficiency tolerance.

Abstract Synergies of transcription factors, chromatin modifiers and their target genes are vital for cell fate determination in human cancer. Although the importance of numerous epigenetic machinery for regulating gliomagenesis has been previously recognized, how chromatin modifiers collaborate with specific transcription factors remains largely elusive. Herein we report that Pontin chromatin remodelling factor acts as a coactivator for LEF1 to activate TGFβ/SMAD signalling, thereby contributing to gliomagenesis. Pontin is highly expressed in gliomas, and its overexpression paralleled the grade elevation and poor prognosis of patients. Functional studies verified its oncogenic roles in GBM cells by facilitating cell proliferation, survival and invasion both in vitro and in vivo. RNA sequencing results revealed that Pontin regulated multiple target genes involved in TGFβ/SMAD signalling. Intriguingly, we found that Pontin amplified TGFβR2 gene transcription by recruiting LEF1, thereby activating TGFβ/SMAD signalling and facilitating gliomagenesis. Furthermore, higher TGFβR2 expression conferred worse patient outcomes in glioma. To conclude, our study revealed that the Pontin-LEF1 module plays a crucial role in driving TGFβR2 gene transcription, which could be exploited to target TGFβ/SMAD signalling for anti-glioma therapy.

Abstract Background In China, nitrogen (N)-deficiency often occurs in Citrus orchards, which is one of the main causes of yield loss and fruit quality decline. Little information is known about the adaptive responses of Citrus carbon (C) and N metabolisms to N-deficiency. Seedlings of ‘Xuegan’ (Citrus sinensis (L.) Osbeck) were supplied with nutrient solution at an N concentration of 0 (N-deficiency), 5, 10, 15 or 20 mM for 10 weeks. Thereafter, we examined the effects of N supply on the levels of C and N in roots, stems and leaves, and the levels of organic acids, nonstructural carbohydrates, NH4 +-N, NO3 −-N, total soluble proteins, free amino acids (FAAs) and derivatives (FAADs), and the activities of key enzymes related to N assimilation and organic acid metabolism in roots and leaves. Results N-deficiency elevated sucrose export from leaves to roots, C and N distributions in roots and C/N ratio in roots, stems and leaves, thus enhancing root dry weight/shoot dry weight ratio and N use efficiency. N-deficient leaves displayed decreased accumulation of starch and total nonstructural carbohydrates (TNC) and increased sucrose/starch ratio as well as a partitioning trend of assimilated C toward to sucrose, but N-deficient roots displayed elevated accumulation of starch and TNC and reduced sucrose/starch ratio as well as a partitioning trend of assimilated C toward to starch. N-deficiency reduced the concentrations of most FAADs and the ratios of total FAADs (TFAADs)/N in leaves and roots. N-deficiency reduced the demand for C skeleton precursors for amino acid biosynthesis, thus lowering TFAADs/C ratio in leaves and roots. N-deficiency increased (decreased) the relative amounts of C-rich (N-rich) FAADs, thus increasing the molar ratio of C/N in TFAADs in leaves and roots. Conclusions Our findings corroborated our hypothesis that C and N metabolisms displayed adaptive responses to N-deficiency in C. sinensis seedlings, and that some differences existed between roots and leaves in N-deficiency-induced alterations of and C and N metabolisms.

The artificial propagation of the endangered Solenaia oleivora, with unknown fish hosts, was performed via in vitro culture with bighead carp, grass carp, common carp, bovine, and rabbit sera. The effects of glochidium density on transformation rates were evaluated, and the development of juveniles that metamorphosed successfully was documented. The control group had a transformation rate of 0 and a contamination rate of 0. No significant differences were found in the transformation and contamination rates of the bighead carp, grass carp, and common carp serum groups, and their transformation rates were significantly higher, and contamination rates were significantly lower than those of the bovine and rabbit serum groups. Moreover, no significant differences were observed in the transformation rates of glochidia (culture density, 2000–5000 glochidia/dish) in contamination-free conditions. Specimen shell length/height increased from 1.08 ± 0.01 on the first day to 3.08 ± 0.29 during the 13th week. On the basis of anterior and posterior growth differences, juvenile growth was divided into the following three phases: the rapid anterior growth period, distinct anterior and posterior idiophase, and rapid posterior growth period. This study not only provides technological support for the artificial propagation of S. oleivora but also lays a foundation for resource recovery.

Objective The aim of this study was to investigate the effects of Enteromorpha powder supplementation on reproduction-related hormones and genes in the late laying period of Zi geese. Methods A total of 312 (1-year-old) Zi geese with similar laying rate were randomly divided into 2 groups with 6 replicates each, each with 21 female geese and 5 male geese. The control group was fed with a basal diet and the test group was fed with a diet containing 3% Enteromorpha powder. The trial period lasted for 7 weeks. Results Our results showed that the laying rate was improved in the test group at each week of trial (p

Abstract Background Primary hyperoxaluria(PH)is a rare autosomal recessive genetic disease that contains three subtypes (PH1, PH2 and PH3). Approximately 80% of PH patients has been reported as subtype PH1, this subtype of PH has been related to a higher risk of renal failure at any age. Several genetic studies indicate that the variants in gene AGXT are responsible for the occurrence of PH1. However, the population heterogeneity of the variants in AGXT makes the genetic diagnosis of PH1 more challenging as it is hard to locate each specific variant. It is valuable to have a complete spectrum of AGXT variants from different population for early diagnosis and clinical treatments of PH1. Case presentation In this study, We performed high-throughput sequencing and genetic analysis of a 6-year-old male PH1 patient from a Chinese family. Two variants (c.346G > A: p.Gly116Arg; c.864G > A: p.Trp288X) of the gene AGXT were identified. We found a nonsense variant (c.864G > A: p.Trp288X) that comes from the proband’s mother and has never been reported previously. The other missense variant (c.346G > A: p.Gly116Arg) was inherited from his father and has been found previously in a domain of aminotransferase, which plays an important role in the function of AGT protein. Furthermore, we searched 110 pathogenic variants of AGXT that have been reported worldwide in healthy local Chinese population, none of these pathogenic variants was detected in the local genomes. Conclusions Our research provides an important diagnosis basis for PH1 on the genetic level by updating the genotype of PH1 and also develops a better understanding of the variants in AGXT by broadening the variation database of AGXT according to the Chinese reference genome.

Abstract Pontin (RUVBL1) is a highly conserved ATPase of the AAA + (ATPases Associated with various cellular Activities) superfamily and is implicated in various biological processes crucial for oncogenesis. Its overexpression is observed in multiple human cancers, whereas the relevance of Pontin to gliomagenesis remains obscure. To gain insights into Pontin involvement in glioma, we performed bioinformatics analyses of Pontin co-expressed genes, Pontin-affected genes, and carried out experimental studies. The results verified that Pontin was upregulated in gliomas. Its higher levels might predict the worse prognosis of glioma patients. The Pontin co-expressed genes were functionally enriched in cell cycle progression and RNA processing. In the nucleus, Pontin promoted cell growth via facilitating cell cycle progression. Using RNA-seq, we found that Pontin knockdown resulted in altered expression of multiple genes, among which the E2F1 targets accounted for a large proportion. Mechanistic studies found that Pontin interacted with E2F1 and markedly amplified the E2F1 transcription response in an ATPase domain-dependent manner. By analyzing the RNA-seq data, we also found that Pontin could impact on the alternative splicing (AS). Both differential expressed genes and AS events affected by Pontin were associated with cell cycle regulation. Taken together, our findings provide novel insights of the importance of Pontin in gliomagenesis by regulating cell cycle and AS, and shed light on the possible application of Pontin as an antineoplastic target in glioma.

The Simultaneous impact of a hollow droplet and a continuous dense droplet on a liquid film was investigated using the coupled level set and volume of fluid (CLSVOF) method. Analyses included fluid dynamics and heat transfer characteristics in impact. Results showed that the interfacial phenomena after impact incorporates spreading, central jet between droplets, edge liquid sheet, and counter jet inside the hollow droplet. The pressure gradient is the major cause for the above phenomena. The significant parameter of impact velocity is closely related to the dynamics and heat transfer for droplets impacting on a liquid film. Droplets with higher impact velocity exhibit a greater spreading factor, central jet height, edge jet height, and counter jet height. Besides, wall heat flux increases more notably for droplets with a higher impact velocity. Compared with the continuous droplet, the hollow droplet shows a smaller spreading factor and edge jet height, a higher wall heat flux, but a narrow thermally affected region. This study provides a fundamental understanding for the application of high-pressure spray combustion.

‘Xuegan’ (Citrus sinensis) seedlings were fertilized 6 times weekly for 24 weeks with 0.5 or 350 μM CuCl2 and 2.5, 10 or 25 μM H3BO3. Cu-toxicity increased Cu uptake per plant (UPP) and Cu concentrations in leaves, stems and roots, decreased water uptake and phosphorus, nitrogen, calcium, magnesium, potassium, sulfur, boron and iron UPP, and increased the ratios of magnesium, potassium, calcium and sulfur UPP to phosphorus UPP and the ratios of leaf magnesium, potassium and calcium concentrations to leaf phosphorus concentration. Many decaying and dead fibrous roots occurred in Cu-toxic seedlings. Cu-toxicity-induced alterations of these parameters and root damage decreased with the increase of boron supply. These results demonstrated that B supplementation lowered Cu uptake and its concentrations in leaves, stems and roots and subsequently alleviated Cu-toxicity-induced damage to root growth and function, thus improving plant nutrient (decreased Cu uptake and efficient maintenance of the other nutrient homeostasis and balance) and water status. Further analysis indicated that the improved nutrition and water status contributed to the boron-mediated amelioration of Cu-toxicity-induced inhibition of seedlings, decline of leaf pigments, large reduction of leaf CO2 assimilation and impairment of leaf photosynthetic electron transport chain revealed by greatly altered chlorophyll a fluorescence (OJIP) transients, reduced maximum quantum yield of primary photochemistry (Fv/Fm), quantum yield for electron transport (ETo/ABS) and total performance index (PIabs,total), and elevated dissipated energy per reaction center (DIo/RC). To conclude, our findings corroborate the hypothesis that B-mediated amelioration of Cu-toxicity involved reduced damage to roots and improved nutrient and water status. Principal component analysis showed that Cu-toxicity-induced changes of above physiological parameters generally decreased with the increase of B supply and that B supply-induced alterations of above physiological parameters was greater in 350 μM Cu-treated than in 0.5 μM Cu-treated seedlings. B and Cu had a significant interactive influence on C. sinensis seedlings.

In this study, a theoretical model of the hot and cold fluid flow, heat transfer, and thermal stress coupling in a baffle plate heat exchanger is established, and numerical simulation is carried out. The flow field in the heat exchanger and the deformation of the heat exchange tube bundles in the baffle plate heat exchanger are studied. The optimization effect of the diversion grid on the flow field at the inlet of the variable section is emphasized. The influence of grid arrangement on the flow field and the performance of the tube bundles are analyzed and discussed. The results show that the backflow situation can be effectively improved and the vortex intensity in the straight section of the cold fluid inlet can be greatly reduced by adding the grid. The deformation of the heat exchange tube bundles illustrates the maximum deformation with 1.39 mm at the position of the strongest backflow. The deformation can be reduced by 26% due to the arrangement of the diversion grid, which is helpful to improve the safety of the device.

Objective To investigate the expression of miRNA⁃449b in gliomas and its effect on cell proliferation and invasion. Methods The expression level of miRNA⁃449b was detected by locked⁃oligonucleotide⁃probe in situ hybridization (ISH) in 60 human glioma specimens of different WHO grades and 10 non⁃tumoral control brain tissues. miRNA⁃449b was quantified by Stem⁃loop quantitation real⁃time polymerase chain reaction (qRT⁃PCR) with U6 as the internal control in U251, LN229 and UC2 cells. EdU staining, MTS method, flow cytometry assay (FCM), Phalloidin staining and Transwell assays were used to evaluate the effect of miRNA⁃449b on the proliferation and invasion of U251 and LN229 cells. Results The expression of miRNA⁃449b was decreased in gliomas compared with that in the non⁃tumoral control brain tissues (P < 0.001, for all), and was significantly declined with the elevation of glioma grades (P < 0.001, for all). miRNA⁃449b level was significantly lower in U251 and LN229 cells than that in UC2 cells (P = 0.001, 0.002). The relative expression of miRNA⁃449b in U251⁃miRNA⁃449b group was higher (P < 0.001), while EdU⁃positive rates (P = 0.029) and cell proliferation (P = 0.004, 0.001, 0.002) was lower than U251⁃Scr group; the relative expression of miRNA⁃449b in LN229⁃miRNA⁃449b group was higher (P < 0.001), while EdU⁃positive rates (P = 0.032) and cell proliferation (P = 0.003, 0.001, 0.004) was lower than LN229⁃Scr group. The percentages of each phase G 0 /G 1 in U251⁃miRNA⁃449b group and LN229⁃miRNA⁃449b group were higher than those in U251 ⁃ Scr group (P < 0.001) and LN229 ⁃ Scr group (P = 0.018) respectively, cell proliferation were lower than U251⁃Scr group (P = 0.002) and LN229⁃Scr group (P = 0.005). F⁃actin was centralized mostly on cell cortex or lamellae in the groups of U251⁃Scr and LN229⁃Scr, whereas diffusely distributed in the groups of U251⁃miRNA⁃449b and LN229⁃miRNA⁃449b. Conclusions miRNA⁃449b is an important tumor⁃suppressive miRNA and its downregulation may be an important cause leading to tumorigenesis of glioblastoma, which could be used as a valuable biomarker to assess the malignant degree of gliomas. Furthermore, the exogenous miRNA⁃449b could effectively inhibit the proliferation and invasion abilities of glioma cells, confirming it has potential value in malignant gliomas therapy. DOI：10.3969/j.issn.1672⁃6731.2019.11.007

Abstract In order to solve the problems of receptor promiscuity and poor blood‐brain barrier (BBB) penetration in the treatment of glioblastomas (GBM), a novel dual‐functional nanocomplex drug delivery system is developed based on the strategy of peptide‐drug conjugates. In this study, SynB3‐PVGLIG‐PTX is designed and screened out by matrix metalloproteinase‐2 (MMP‐2), to which it exhibits the best affinity. The MMP‐2‐sensitive peptide (PVGLIG) and a cell‐penetration peptide (SynB3) are combined to form a dual‐functional peptide. Moreover, as a drug‐peptide nanocomplex, SynB3‐PVGLIG‐PTX exhibited a high potential to form an aggregation with good solubility that can release paclitaxel (PTX) through the cleavage of MMP‐2. From a functional perspective, it is found that SynB3‐PVGLIG‐PTX can specifically inhibit the proliferation, migration, and invasion of GBM cells in vitro in the presence of MMP‐2, in contrast to that observed in MMP‐2 siRNA transfected cells. Further investigation in vivo shows that SynB3‐PVGLIG‐PTX easily enters the brain of U87MG xenograft nude mice and can generate a better suppressive effect on GBM through a controlled release of PTX from SynB3‐PVGLIG‐PTX compared with PTX and temozolomide. Thus, it is proposed that SynB3‐PVGLIG‐PTX can be used as a novel drug‐loading delivery system to treat GBM due to its specificity and BBB permeability.

Purpose: To investigate the potential association between lifestyles, including cigarette smoking, alcohol consumption, and physical exercise at the time of biopsy and the risk for developing end-stage renal failure (ESRF) among IgA nephropathy (IgAN) patients within 10 years. Methods: A case–control study was carried out. Seventy-seven ESRF patients with the primary cause of IgAN were enrolled as cases. Seventy-seven IgAN patients who had not progressed to ESRF after being diagnosed for over 10 years served as controls. Smoking, alcohol consumption and physical exercise related data and baseline clinical features were collected from their medical records and confirmed by phone calls. Results: The case group had higher proportions of males, smokers, drinkers, and physical inactivity individuals than the controls had. Alcohol drinking history (/1 year, OR 1.32, p

Membranes are widely used for liquid separations such as removing solute components from solvents or liquid/liquid separations. Due to negligible vapor pressure, adjustable physical properties, and thermal stability, the application of ionic liquids (ILs) has been extended to fabricating a myriad of membranes for liquid separations. A comprehensive overview of the recent developments in ILs in fabricating membranes for liquid separations is highlighted in this review article. Four major functions of ILs are discussed in detail, including their usage as (i) raw membrane materials, (ii) physical additives, (iii) chemical modifiers, and (iv) solvents. Meanwhile, the applications of IL assisted membranes are discussed, highlighting the issues, challenges, and future perspectives of these IL assisted membranes in liquid separations.

The basal media M199 or MEM was utilized in the classical method of vitro culture of glochidia where 1-5% CO2 was required to maintain stable physiological pH for completion of non-parasitic metamorphosis. The classical method encounters a great challenge to those glochidia which undergo development of visceral tissue but significantly increase in size during metamorphosis. The improved in vitro culture techniques and classical methods were firstly compared for non-parasitic metamorphosis and development of glochidia in pink heelsplitter. Based on the improved method, the optimal vitro culture media was further selected from 14 plasmas or sera, realizing the non-parasitic metamorphosis of axe-head glochidia for the first time. The results showed that addition of different plasma (serum) had significant effect on glochidial metamorphosis in pink heelsplitter. Only glochidia in the skewband grunt and red drum groups could complete metamorphosis, the metamorphosis rate in skewband grunt was 93.3±3.1% at 24±0.5°C, significantly higher than in marine and desalinated red drum. Heat-inactivated treatment on the plasma of yellow catfish and Barbus capito had significant effect on glochidia survival and shell growth. The metamorphosis rate also varied among different gravid period, and generally decreased with gravid time. Further comparison of free amino acid and fatty acid indicated that the taurine of high concentration was the only amino acid that might promote the rapid growth of glochidial shell, and the lack of adequate DPA and DHA might be an important reason leading to the abnormal foot and visceral development. Combined with our results of artificial selection of host fish, we tentatively established the mechanism of its host specialists in pink heelsplitter for the first time. This is the first report on non-parasite metamorphosis of axe-head glochidia based on our improved vitro culture method, which should provide important reference to fundamental theory research of glochidia metamorphosis and also benefit for better understand of mechanism of host specialists and generalists of Unionidae species.

The main role of cold water discharged from ocean thermal energy conversion (OTEC) systems is that deep ocean water, which is dense and nutrient-rich, is released through the condenser and discharged into the ocean surface. We present a numerical simulation in which a cold-water is discharged into a temperature-stratified fluid. A semi-empirical formula relating the outlet flow and the plunging depth was obtained by model analysis, and the k multiplier was 1.15. The model results are consistent with the experimental data.

Doubly uniparental inheritance (DUI) of mitochondrial DNA, found only in some bivalve families and characterized by the existence of gender-associated mtDNA lineages that are inherited through males (M-type) or females (F-type), is one of the very few exceptions to the general rule of strict maternal mtDNA inheritance in animals. M-type sequences are often undetected and hence still underrepresented in the GenBank, which hinders the progress of the understanding of the DUI phenomenon. We have sequenced and analyzed the complete M and F mitogenomes of a freshwater mussel, Potamilus alatus. The M-type was 493 bp longer (M = 16 560, F = 16 067 bp). Gene contents, order and the distribution of genes between L and H strands were typical for unionid mussels. Candidates for the two ORFan genes (forf and morf) were found in respective mitogenomes. Both mitogenomes had a very similar A+T bias: F = 61% and M = 62.2%. The M mitogenome-specific cox2 extension (144 bp) is much shorter than in other sequenced unionid mitogenomes (531-576 bp), which might be characteristic for the Potamilus genus. The overall topology of the phylogenetic tree is in very good agreement with the currently accepted phylogenetic relationships within the Unionidae: both studied sequences were placed within the Ambleminae subfamily clusters in the corresponding M and F clades.

In the original version of Figure 6P, the labels for AAV-EGFP and AAV-Cre were inverted. The correct figure part is below. The HTML and PDF files have been updated online. [...]