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As a member of the forkhead box L gene family, foxl2 plays a significant role in gonadal development and the regulation of reproduction. During the evolution of deuterostome, whole genome duplication (WGD)-enriched lineage diversifications and regulation mechanisms occurs. However, only limited research exists on foxl2 duplication in teleost or other vertebrate species. In this study, two foxl2 paralogs, foxl2 and foxl2l , were identified in the transcriptome of spotted knifejaw ( Oplegnathus punctatus ), which had varying expressions in the gonads. The foxl2 was expressed higher in the ovary, while foxl2l was expressed higher in the testis. Phylogenetic reconstruction, synteny analysis, and the molecular evolution test confirmed that foxl2 and foxl2l likely originated from the first two WGD. The expression patterns test using qRT-PCR and ISH as well as motif scan analysis revealed evidence of potentially functional divergence between the foxl2 and foxl2l paralogs in spotted knifejaw. Our results indicate that foxl2 and foxl2l may originate from the first two WGD, be active in transcription, and have undergone functional divergence. These results shed new light on the evolutionary trajectories of foxl2 and foxl2l and highlights the need for further detailed functional analysis of these two duplicated paralogs.

The quality of Schisandra chinensis (Turcz.) Baill. ( S. chinensis ) is principally attributed to lignan compounds. In this paper, a simple and rapid strategy for simultaneous extraction and determination of 10 lignans from S. chinensis was established through matrix solid-phase dispersion (MSPD) assisted by diol-functionalized silica (Diol). The experimental parameters for MSPD extraction were screened using the response surface methodology (RSM). Diol (800 mg) was used as a dispersant and methanol (MeOH, 85%, v / v ) as an eluting solvent (10 mL), resulting in a high extraction efficiency. MSPD extraction facilitated the combination of extraction and purification in a single step, which was less time-consuming than and avoided the thermal treatment involved in traditional methods. The simultaneous qualification and quantification of 10 lignans was achieved by combining MSPD and high-performance liquid chromatography (HPLC). The proposed method offered good linearity and a low limit of detection starting from 0.04 (schisandrin C) to 0.43 μg/mL (schisantherin B) for lignans, and the relative standard deviation (RSD, %) values of precision were acceptable, with a maximum value of 1.15% (schisantherin B and schisanhenol). The methodology was successfully utilized to analyze 13 batches of S. chinensis from different cultivated areas of China, which proved its accuracy and practicability in the quantitative analysis of the quality control of S. chinensis .

Alzheimer's disease (AD) is a critical health issue worldwide that has a negative impact on patients' quality of life, as well as on caregivers, society, and the environment. Positron emission tomography (PET)/computed tomography (CT) and neuropsychological scales can be used to identify AD and mild cognitive impairment (MCI) early, provide a differential diagnosis, and offer early therapies to impede the course of the illness. However, there are few reports of large-scale 11 C-PIB-PET/CT investigations that focus on the pathology of AD and MCI. Therefore, further research is needed to determine how neuropsychological test scales and PET/CT measurements of disease progression interact., 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 © 2023 Zhao, Du, Chen, Zhang and Xu.)

The multi-enzyme coupling reaction system has become a promising biomanufacturing platform for biochemical production. Tyr is an essential amino acid, but the limited solubility restricts its use. Tyrosyl dipeptide has been paid more attention due to its higher solubility. In this study, an efficient enzymatic cascade of Ala-Tyr synthesis was developed by a L-amino acid ligase together with polyphosphate kinase (PPK). Two L-amino acid ligases from Bacillus subtilis and Bacillus pumilus were selected and applied for Ala-Tyr synthesis. The L-amino acid ligase from B. subtilis (Bs) was selected and coupled with the PPK from Sulfurovum lithotrophicum (PPKSL) for regenerating ATP to produce Ala-Tyr in one pot. In the optimization system, 40.1 mM Ala-Tyr was produced within 3 h due to efficient ATP regeneration with hexametaphosphate (PolyP(6)) as the phosphate donor. The molar yield was 0.89 mol/mol based on the substrates added, while the productivity of Ala-Tyr achieved 13.4 mM/h, which were the highest yield and productivity ever reported about Ala-Tyr synthesis with L-amino acid ligase., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Digestates from different anaerobic digesters are promising substrates for microalgal culture, leading to effective wastewater treatment and the production of microalgal biomass. However, further detailed research is needed before they can be used on a large scale. The aims of this study were to investigate the culture of Chlorella sp. in Digestate M from anaerobic fermentation of brewer's grains and brewery wastewater (BWW) and to explore the potential use of the biomass produced under different experimental conditions, including diverse cultivation modes and dilution ratios. Cultivation in Digestate M initiated from 10% (v/v) loading, with 20% BWW, obtained maximum biomass production, reaching 1.36 g L -1 that was 0.27g L -1 higher than 1.09 g L -1 of BG11. In terms of Digestate M remediation, the maximum removal of ammonia nitrogen (NH 4 + -N), chemical oxygen demand, total nitrogen, and total phosphorus reached 98.20%, 89.98%, 86.98%, and 71.86%, respectively. The maximum lipid, carbohydrate, and protein contents were 41.60%, 32.44%, and 27.72%, respectively. The growth of Chlorella sp. may be inhibited when the Y(II)-F v /F m ratio is less than 0.4., 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 © 2023 Wang, Zhao, Yu, Du, Zhang, Sun and Song.)

Gibberellic acids had been proven to improve the fruit quality and storability by delaying deterioration and maintaining the antioxidant system. In this study, the effect of GA 3 spraying at different concentrations (10, 20, and 50 mg L -1 ) on the quality of on-tree preserved 'Shixia' longan was examined. Only 50 mg L -1 GA 3 significantly delayed the decline of soluble solids (22.0% higher than the control) and resulted in higher total phenolics content (TPC), total flavonoid content (TFC), and phenylalanine ammonia-lyase activity in pulp at the later stages. The widely targeted metabolome analysis showed that the treatment reprogrammed secondary metabolites and up-regulated many tannins, phenolic acids, and lignans during the on-tree preservation. More importantly, the preharvest 50 mg L -1 GA 3 spraying (at 85 and 95 days after flowering) led to significantly delayed pericarp browning and aril breakdown, as well as lower pericarp relative conductivity and mass loss at the later stages of room-temperature storage. The treatment also resulted in higher antioxidants in pulp (vitamin C, phenolics, and reduced glutathione) and pericarp (vitamin C, flavonoids, and phenolics). Therefore, preharvest 50 mg L -1 GA 3 spraying is an effective method for maintaining the quality and up-regulating antioxidants of longan fruit during both on-tree preservation and room-temperature storage.

Training of viewing an altered-reality environment dichoptically has been found to reactivate human adult ocular dominance plasticity, allowing improvement of vision for amblyopia. One suspected mechanism for this training effect is ocular dominance rebalancing through interocular disinhibition. Here, we investigated whether the training modulated the neural responses reflecting interocular inhibition. Thirteen patients with amblyopia and 11 healthy controls participated in this study. Before and after six daily altered-reality training sessions, participants watched flickering video stimuli with their steady-state visually evoked potential (SSVEP) signals recorded simultaneously. We assessed the amplitude of SSVEP response at intermodulation frequencies, which was a potential neural indicator of interocular suppression. The results showed that training weakened the intermodulation response only in the amblyopic group, which was in agreement with the hypothesis that the training reduced interocular suppression specific to amblyopia. Moreover, even one month after the training ended, we could still observe this neural training effect. These findings provide preliminary neural evidence in support of the disinhibition account for treating amblyopia. We also explain these results with the ocular opponency model, which, to our knowledge, is the first time for this binocular rivalry model to be used in explaining long-term ocular dominance plasticity., (© 2023 New York Academy of Sciences.)

As the main flavor components of tea, the contents of epigallocatechin-3-gallate (EGCG), theanine and caffeine are regulated by ambient temperature. However, whether the biosynthesis of EGCG, theanine and caffeine in response to temperature is regulated by endogenous hormones and its mechanism is still unclear. In this study, tea cuttings cultivated in the phytotron which treated at different temperatures 15℃, 20℃, 25℃ and 30℃, respectively. The UPLC and ESI-HPLC-MS/MS were used to determine the contents of EGCG, theanine, caffeine and the contents of phytohormones in one leaf and a bud. The results showed that indoleacetic acid (IAA), gibberellin 1(GA1) and gibberellin 3 (GA3) were significantly correlated with the content of EGCG; Jasmonic acid (JA), jasmonate-isoleucine (JA-Ile) and methyl jasmonate (MeJA) were strongly correlated with theanine content; IAA, GA1 and gibberellin 4 (GA4) were significantly correlated with caffeine content at different temperatures. In order to explore the internal intricate relationships between the biosynthesis of these three main taste components, endogenous hormones, and structural genes in tea plants, we used multi-omics and multidimensional correlation analysis to speculate the regulatory mechanisms: IAA, GA1 and GA3 up-regulated the expressions of chalcone synthase ( CsCHS ) and trans-cinnamate 4-monooxygenase ( CsC4H ) mediated by the signal transduction factors auxin-responsive protein IAA ( CsIAA ) and DELLA protein ( CsDELLA ), respectively, which promoted the biosynthesis of EGCG; IAA, GA3 and GA1 up-regulated the expression of CsCHS and anthocyanidin synthase ( CsANS ) mediated by CsIAA and CsDELLA , respectively, via the transcription factor WRKY DNA-binding protein ( CsWRKY ), and promoted the biosynthesis of EGCG; JA, JA-Ile and MeJA jointly up-regulated the expression of carbonic anhydrase ( CsCA ) and down-regulated the expression of glutamate decarboxylase ( CsgadB ) mediated by the signal transduction factors jasmonate ZIM domain-containing protein ( CsJAZ ), and promoted the biosynthesis of theanine; JA, JA-Ile and MeJA also jointly inhibited the expression of CsgadB mediated by CsJAZ via the transcription factor CsWRKY and AP2 family protein ( CsAP2 ), which promoted the biosynthesis of theanine; IAA inhibited the expression of adenylosuccinate synthase ( CspurA ) mediated by CsIAA via the transcription factor CsWRKY ; GA1 and gibberellin 4 (GA4) inhibited the expression of CspurA mediated by CsDELLA through the transcription factor CsWRKY , which promoted the biosynthesis of caffeine. In conclusion, we revealed the underlying mechanism of the biosynthesis of the main taste components in tea plant in response to temperature was mediated by hormone signal transduction factors, which provided novel insights into improving the quality of tea., Competing Interests: Author LJW was employed by Fujian Sanan Sino-Science Photobiotech Co., Ltd. The remaining 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 © 2023 Zhu, Liu, Lu, Du, Xiang, Cheng, Tan, Huang, Wu, Kong, Shi, Wu and Lin.)

Bisphenol A (BPA) can induce complex regulatory mechanisms in many aquatic organisms, and it is difficult to find a suitable analytical method to efficiently enrich key genes responding to BPA exposure. In this study, zebrafish embryo transcriptomic data were obtained from two types of different BPA exposure methods. After BPA exposure, three differential gene enrichment methods were used jointly to identify up-regulated genes or pathways in zebrafish embryo larvae. The results showed that the systemic lupus erythematosus signaling pathway was significantly enriched in all BPA exposure groups. It was also noteworthy that most of the up-regulated genes in systemic lupus erythematosus signaling were histones. In conclusion, this study suggested that autoimmunity signaling was the most common important pathway in zebrafish embryo-larvae response to different BPA exposures, and histones may play a key role in response to low-concentration BPA., 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 © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: To assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies., Methods: 10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13., Results: The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly., Conclusion: The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.

Background: A functional artificial ovary is a promising strategy to recover fertility and restore endocrine function in cancer patients. The aim of this study is to optimize the follicle isolation protocol for cryopreserved human ovarian tissues. Methods: Each of the cryopreserved human ovarian cortex pieces (OCPs) from 10 patients was cut into two equal parts and randomly distributed into two treatment groups. Group 1: OCPs digested with Tumor Dissociation Enzyme (TDE); Group 2: OCPs digested with Liberase Dispase High (DH). The efficiency of both groups were evaluated in terms of yield, viability, morphology, and a short-term in vitro culture (IVC) in alginate scaffolds. Results: The TDE can isolate more primordial follicles and smaller diameter of follicles than Liberase DH. The TDE also enabled the isolation of more bright red follicles, higher percent of viable follicles, more morphologically normal follicles, and lower oxidative stress levels compared with Liberase DH. After eight days of IVC, follicles in the TDE group had a higher growth rate from Day 0 to Day 8, and higher viability on Day 8 than the Liberase DH Group. Conclusion: The TDE can be considered an alternative to Liberase DH, enables the isolation of a higher number of healthy follicles from human OCPs, and improves follicle survival after IVC in contrast to Liberase DH.

Although the effects of phytohormones (mainly salicylic acid) on the storability of longan fruit have been reported, the relationship between postharvest hormone variation and signal transduction and storability remains unexplored. The basis of physiology, biochemistry, hormone content and signalling for the storability difference at room-temperature between 'Shixia' and 'Luosanmu' longan fruit were examined. 'Luosanmu' longan exhibited faster pericarp browning, aril breakdown and rotting during storage. 'Luosanmu' pericarp exhibited higher malondialdehyde but faster decreased total phenolics, flavonoid, glutathione, vitamin C, catalase activity and gene expression. Higher H 2 O 2 and malondialdehyde but lower glutathione, glutathione-reductase and peroxidase activities, while higher activities and gene expressions of polygalacturonase, β-galactosidase and cellulose, lower covalent-soluble pectin, cellulose and hemicellulose but higher water-soluble pectin were observed in 'Luosanmu' aril. Lower abscisic acid and methyl jasmonate but higher expressions of LOX2 , JAZ and NPR1 in pericarp, while higher abscisic acid, methyl jasmonate and salicylic acid together with higher expressions of ABF , JAZ , NPR1 and PR-1 in 'Luosanmu' aril were observed. In conclusion, the imbalance between the accumulation and scavenging of active oxygen in 'Luosanmu' longan might induce faster lipid peroxidation and senescence-related hormone signalling and further the polymerization of phenolics in pericarp and polysaccharide degradation in aril.

Androstenedione (ADSD) was the main androgen detected in wastewaters. Chlorella was the most widely used plant in biological wastewater treatment process. In order to understand the toxicological response of chlorella to ADSD contamination, we used the weighted gene co-expression network analysis (WGCNA) method to systematically analyze the gene regulatory networks of chlorella after ADSD treatments. Total of 25 modules was identified from gene co-expression networks, and the turquoise module were selected for GO and KEGG enrichment analysis. Results showed that most hub genes were associated with chloroplast organizations or photosystems processes. Among them, the expressions profiles of hcar, nol, pao and sgr genes were highly correlated to the content fluctuations of chlorophylls after different ADSD treatments. All these results demonstrated that chlorophylls play a key role in preventing cell damage of chlorella caused by ADSD contamination. Besides, we proposed a possible chlorophyll metabolism pathway in chlorella response to ADSD contamination., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Cytosolic Ca2+ is a highly dynamic, tightly regulated and broadly conserved cellular signal. Ca2+ dynamics have been studied widely in cellular monocultures, yet organs in vivo comprise heterogeneous populations of stem and differentiated cells. Here, we examine Ca2+ dynamics in the adult Drosophila intestine, a self-renewing epithelial organ in which stem cells continuously produce daughters that differentiate into either enteroendocrine cells or enterocytes. Live imaging of whole organs ex vivo reveals that stem-cell daughters adopt strikingly distinct patterns of Ca2+ oscillations after differentiation: enteroendocrine cells exhibit single-cell Ca2+ oscillations, whereas enterocytes exhibit rhythmic, long-range Ca2+ waves. These multicellular waves do not propagate through immature progenitors (stem cells and enteroblasts), of which the oscillation frequency is approximately half that of enteroendocrine cells. Organ-scale inhibition of gap junctions eliminates Ca2+ oscillations in all cell types - even, intriguingly, in progenitor and enteroendocrine cells that are surrounded only by enterocytes. Our findings establish that cells adopt fate-specific modes of Ca2+ dynamics as they terminally differentiate and reveal that the oscillatory dynamics of different cell types in a single, coherent epithelium are paced independently., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Background: Abnormalities of lymphocyte subsets and cytokine profiles have been observed in most patients with coronavirus disease (COVID-19). Here, we explore the role of lymphocyte subsets and cytokines on hospital admission in predicting the severity of COVID-19., Methods: This study included 214 patients with COVID-19 who were treated at Chongqing University Three Gorges Hospital from January 19, 2020 to April 30, 2020. Any mutants were not detected in the studied patients. Patients were divided into non-intensive care unit (ICU) (mild/moderate) group and ICU (severe/critical) group, according to the severity of the disease. Clinical and laboratory data, including peripheral lymphocyte subsets and cytokines, were analyzed and compared. Logistic regression was used to analyze the predictive factors for ICU admission. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of selected indicators for the severity of COVID-19., Results: Of the 214 patients enrolled, 161 were non-ICU patients and 53 were ICU patients. Lymphopenia was observed in nearly all of ICU patients (96.2%) and 84.5% of non-ICU patients on hospital admission. The absolute number of lymphocytes, CD3 + T cells, CD4 + T cells, CD8 + T cells, CD19 + B cells, and natural killer (NK) cells were lower in ICU group (659.00 × 10 6 /L, 417.00 × 10 6 /L, 261.00 × 10 6 /L, 140.00 × 10 6 /L, 109.00 × 10 6 /L, 102.00 × 10 6 /L, respectively) than in non-ICU group (1063.00 × 10 9 /L, 717.00 × 10 6 /L, 432.00 × 10 6 /L, 271.00 × 10 6 /L, 133.00 × 10 6 /L, 143.00 × 10 6 /L, respectively). Interleukin (IL)-6 was significantly higher in ICU patients than in non-ICU patients (18.08 pg/mL vs. 3.13 pg/mL, P < 0.001). Multivariate logistic regression analysis showed that age (odds ratio: 1.067 [1.034-1.101]), diabetes mellitus (odds ratio: 9.154 [2.710-30.926]), CD3 + T cells (odds ratio: 0.996 [0.994-0.997]), and IL-6 (odds ratio: 1.006 [1.000-1.013]) were independent predictors for the development of severe disease. ROC curve analysis showed that the area under the ROC curve (AUC) of CD3 + T cells and IL-6 was 0.806 (0.737-0.874) and 0.785 (0.705-0.864), respectively, and the cutoff values were 510.50 × 10 6 /L (sensitivity, 71.7%; specificity, 79.5%) and 6.58 pg/mL (77.4%, 74.5%), respectively. There were no statistical differences among all tested indicators of lymphocyte subsets and cytokines between severe group (n = 38) and critical group (n = 15) on hospital admission or ICU admission, respectively., Conclusions: The levels of lymphocyte subsets decreased and the level of IL-6 increased significantly in ICU COVID-19 patients compared with non-ICU COVID-19 patients. Therefore, the number of CD3 + T cells and the level of IL-6 on hospital admission may serve as predictive factors for identifying patients with wild-type virus infection who will have severe disease., (© 2022. The Author(s).)

Objective: The aim of the present study was to evaluate the clinical value of comprehensive nursing intervention in preventing ventilator-associated pneumonia (VAP)., Methods: Eligible 116 patients who received mechanical ventilation in our hospital from June 2018 to December 2019 were equally randomized into a control group and an observation group. The observation group adopted comprehensive nursing while conventional nursing was adopted in the control group accordingly. The mechanical ventilation time, hospitalization time, VAP incidence, psychological status, quality of life and nursing satisfaction were compared., Results: The mechanical ventilation and hospitalization time in the observation group was shorter compared to that in the control group ( P < 0.05). For the VAP incidence, the observation group exhibited a significantly lower level than the control group (3.44% vs 13.79%) ( P < 0.05). SAS and SDS scores before intervention were not different between the groups ( P > 0.05); after intervention, the scores in both groups decreased, and the scores of the former group were significantly lower than those of the latter group ( P < 0.05). The quality of life scores before intervention did not show any significant difference ( P > 0.05); after intervention, a significant increase of quality of life in the observation group was observed compared to the control group (P < 0.05). The nursing satisfaction rate in the observation group was found to be higher when compared with that in the control group (94.87% vs 81.03%, P < 0.05). After intervention, the oxygenation index, the ratio of alveolar arterial oxygen partial pressure difference to arterial oxygen partial pressure (P(A-a)DO 2 /PaO 2 ), and oxygen partial pressure in both groups of patients were improved compared with those before intervention. After intervention, a significant increase in the oxygen partial pressure and oxygenation index was identified in the observation group compared with the control group, while the P(A-a)DO 2 /PaO 2 was found to be in a lower level (P < 0.05)., Conclusion: Comprehensive nursing intervention can shorten the mechanical ventilation and hospitalization time, prevent the occurrence of VAP, ease the negative emotions of patients, and improve their quality of life as well as nursing satisfaction., Competing Interests: None., (AJTR Copyright © 2021.)

Background: Ovarian tissue cryopreservation has a wide range of cancerous indications. Avoiding relapse becomes a specific concern that clinicians frequently encounter. The data about the comparative viability of cancer cells after cryopreservation are limited. This study aimed to evaluate the effect of cryopreservation on breast cancer cells., Methods: We used in-vitro cultured ZR-75-1 and MDA-MB-231 cell lines. Cell samples of each lineage were distributed into the non-intervened and cryopreserved groups. The cryopreservation procedures comprised programmed slow freezing followed by thawing at 100 °C, 60 s. Biological phenotypes and the related protein markers were compared between the two groups. The EVOS FL Auto 2 Cell Image System was used to monitor cell morphology. Cell proliferation, motility, and penetration were characterized by CCK-8, wound-healing, and transmembrane assay, respectively. The expression of Ki-67, P53, GATA3, E-cadherin, Vimentin, and F-Actin was captured by immunofluorescent staining and western blotting as the proxy measurements of the related properties. The chorioallantoic membrane (CAM) xenotransplantation was conducted to explore angiogenesis induced by cancer cells., Results: After 5 days in vitro culture, the cell concentration of cryopreserved and non-intervened groups was 15.7 × 10 4 vs. 14.4 × 10 4 cells/ml, (ZR-75-1, p > 0.05), and 25.1 × 10 4 vs. 26.6 × 10 4 cells/ml (MDA-MB-231, p > 0.05). Some cryopreserved ZR-75-1 cells presented spindle shape with filopodia and lamellipodia and dissociated from the cell cluster after cryopreservation. Both cell lines demonstrated increased cell migrating capability and invasion after cryopreservation. The expression of Ki-67 and P53 did not differ between the cryopreserved and non-intervened groups. E-cadherin and GATA3 expression downregulated in the cryopreserved ZR-75-1 cells. Vimentin and F-actin exhibited an upregulated level in cryopreserved ZR-75-1 and MDA-MB-231 cells. The cryopreserved MDA-MB-231 cells induced significant angiogenesis around the grafts on CAM with the vascular density 0.313 ± 0.03 and 0.342 ± 0.04, compared with that of non-intervened cells of 0.238 ± 0.05 and 0.244 ± 0.03, p < 0.0001., Conclusions: Cryopreservation promotes breast cancer cells in terms of epithelial-mesenchymal transition and angiogenesis induction, thus increasing metastasis risk.

Objective: To comprehensively understand the basic construction of intensive care unit (ICU) in the secondary and tertiary hospitals in Xinjiang Uygur Autonomous Region, and to provide a theoretical basis for the development direction of critical care medicine and the rational allocation of medical resources in our region., Methods: On the March 14th, 2020, a cross-sectional survey of 147 ICU in 122 hospitals in Xinjiang Uygur Autonomous Region was conducted using an online questionnaire. The survey included 6 modules: the basic conditions of the hospital, ICU profile, ICU human resources status, equipment allocation, technology development, and ICU quality control., Results: Among the 147 ICUs, there were 69 ICUs in tertiary hospital and 78 ICUs in secondary hospital. 75.51% (111/147) were comprehensive ICU and 24.49% (36/147) were specialized ICU. The total number of ICU beds was 1 818, accounted about 2.43% (1 818/74 912) of the total number of hospital beds. In ICU terms of human resourse, physicians/beds ratio was 0.54:1, and nurses/beds ratio was 1.55:1. Physicians/beds ratio in the secondary hospitals was 0.52:1, and nurses/beds was 1.45:1; physicians/beds ratio in the tertiary hospital was 0.56:1, and nurses/beds ratio was 1.79:1. The ICU management model was mainly closed management (82.99%, 122/147), and the proportion of closed management in tertiary hospitals was 88.41% (61/69), which was higher than that in secondary hospitals (78.21%, 61/78). In aspect of ICU equipment, the invasive ventilator/bed ratio, enteral nutrition infusion pump/bed ratio, and blood purifier/bed ratio in the tertiary hospitals were significantly higher than those in the secondary hospitals [0.70 (0.46, 1.00) vs. 0.45 (0.33, 0.67), 0.18 (0.00, 0.56) vs. 0.00 (0.00, 0.13), 0.08 (0.00, 0.13) vs. 0.00 (0.00, 0.10), respectively, all P < 0.01]. In the tertiary hospital, the chest sputum excretion device, blood gas analyzer, blood purification instrument, transport ventilator, fiber bronchoscope, enteral nutrition infusion pump, bedside ultrasound machine, continuous hemodynamics and oxygen metabolism monitor, electroencephalogram bispectral index monitor, bedside electroencephalography machine and extracorporeal membrane oxygenation (ECMO) were also superior to the secondary hospitals. ICU technologies, such as deep venipuncture, jejunal nutrition tube placement, percutaneous tracheotomy, invasive blood pressure monitoring, invasive hemodynamic monitoring, bedside ultrasound examination, continuous blood purification, fiber bronchoscopy, high frequency ventilation, intra-aortic balloon counterpulsation (IABP), and ECMO had also performed better than secondary hospitals. In the management of ICU medical quality control, in tertiary hospitals, the proportions of single or isolated room for patients with drug-resistant bacteria, 1-hour bundle and hemodynamic monitoring for patients with septic shock, routine prone position ventilation and lung recruitment for patients with acute respiratory distress syndrome (ARDS), common analgesic, and use of personal digital assistant (PDA) for pre-operation scan code by nurses and electronic medical record for routine rounds were significantly higher than those in secondary hospitals (91.30% vs. 85.90%, 68.12% vs. 48.72%, 85.51% vs. 70.51%, 28.99% vs. 12.82%, 85.51% vs. 61.54%, 76.81% vs. 61.54%, 71.01% vs. 29.49, 49.28% vs. 28.21%, respectively), and the differences were statistically significant (all P < 0.05). 89.74% (70/78) ICU in secondary hospitals and 89.86% (62/69) of tertiary hospitals used acute physiology and chronic health evaluation II (APACHE II) to evaluate the severity of critically ill patients; in terms of sequential organ failure assessment (SOFA), the difference between the secondary hospitals and the tertiary hospitals was not statistically significant (51.28% vs. 62.32%, χ 2 = 1.814, P = 0.178)., Conclusions: Although the ICU construction of the tertiary hospitals in Xinjiang Uygur Autonomous Region is more complete than secondary hospitals, there is a big gap between the requirements of the national guidelines and the developed regions in the east. The ICU's investment in human resource, equipment and supporting facilities allocation, promotion of suitable technology, and medical quality control management should be increased to promote the development of critical care medicine in Xinjiang Uygur Autonomous Region.

Auto-transplantation of cryopreserved ovarian tissue for cancer survivors comes with the primary concern of the possible existence of cancer cells in the transplanting tissue. Lineal cancer cells are presented by industry in form of separated cells (suspensions). Data about experimental models for evaluation of a cryopreservation effect on viability of compacted lineal cancer cells is currently limited. This study aims to develop a suitable experimental model for cryobiological investigations of compacted cancer cells obtained after in vitro culture of a cell suspension. Suspended lineal breast cancer cells (ZR-75-1 and MDA-MB-231) were in vitro cultured in AIM V medium for formation of monolayer. Evaluation of the cell viability was performed by healing assay, transmembrane cell migration, invasion assay and immunofluorescent test of F-actin. It was established the possibility of formation of monolayer from viable cancer cells, scarification of monolayer of these cells and formation of compacted fragments. It is described also a behaviour of compacted cells during cryopreservation (saturation by permeable cryoprotectants, thawing and removal of cryoprotectants). The described method can be used for cryobiological investigations of lineal suspended cancer cells in compacted form as a model of tissues contaminated by malignant cells and solid tumors.

Objective: To investigate the effect of terlipressin on prognosis of adult septic shock patients., Methods: All randomized controlled clinical trials (RCT) of terlipressin in the treatment of adult septic shock patients from January 1980 to December 2019 were retrieved from CNKI, Wanfang, SinoMed, PubMed, Embase, Springer Link, Cochrane Library, Google Scholar, and etc. Patients in the treatment group received terlipressin while patients in the control group received norepinephrine or other vasopressors. Main outcome indicator was mortality. Secondary outcome indicators included the incidence of severe adverse events, limb peripheral ischemic events and renal complications. Literature screening, data extraction and quality evaluation were conducted by two researchers respectively. Meta-analysis was performed with RevMan 5.3 software. Funnel plot was used to analyze the publication bias., Results: A total of 507 related literatures were retrieved. According to the inclusion and exclusion criteria, 8 RCT studies were finally included, with a total of 811 patients. One study was considered to have a lower risk of bias, 6 studies had uncertain risk of bias, and 1 study had a higher risk of bias. The Meta-analysis showed that terlipressin did not significantly improve the mortality of septic shock patients compared with the control group [odds ratio (OR) = 0.89, 95% confidence interval (95%CI) was 0.67-1.19, P = 0.45]; increased the incidence of severe adverse events (OR = 2.98, 95%CI was 1.99-4.45, P < 0.000 01); there was a tendency to increase the incidence of limb peripheral ischemic events, but without statistical difference (OR = 10.81, 95%CI was 0.88-133.19, P = 0.06); and reduced the incidence of renal complications (OR = 0.30, 95%CI was 0.09-0.96, P = 0.04). Funnel plot analysis indicated that there might be publication bias in a study on case fatality and incidence of serious adverse events in the included literature. No significant publication bias was found in studies on the incidence of limb peripheral ischemic events and the incidence of kidney-related complications., Conclusions: The available evidence suggests that terlipressin could not significantly improve mortality in adult's septic shock patients, but it may reduce the incidence of renal complications. A tendency to increase the incidence of limb peripheral ischemic events in the terlipressin-treated group needs to be emphasized.

Objective: Sepsis is one of the leading causes of inpatient deaths worldwide and is a major challenge in clinical work. The diagnosis and treatment of sepsis has been a research hotspot in critical care medicine. Through the efforts during the past decades, deeper understanding of the pathophysiology of sepsis and progress in the treatment have been made. But the latest sepsis guidelines and bundle strategies remain controversial, and clinical researches on sepsis are mixed. So far, there is no specific therapy for sepsis, and the mortality is still very high. Important researches and viewpoints in the field of sepsis in recent years are summarized and analyzed, so as to provide reference for the treatment of sepsis patients.

Objective: To systematically evaluate the effect of vitamin C on prognosis of critically ill patients., Methods: Randomized controlled trials (RCT) about vitamin C treatment for critically ill patients were searched in CNKI, CBM, VIP database, Wanfang database, PubMed, Springer Link, Embase, Web of Science, and Cochrane Library from their inception to May 2019. Patients in the treatment group received ascorbic acid while patients in the control group received placebo or other treatment. Outcome measures included mortality, the length of intensive care unit (ICU) stay, the length of hospital stay, and incidence of atrial fibrillation. Two researchers were responsible for literature screening, data extraction and quality evaluation independently. Meta-analysis was performed with RevMan 5.2 software. The publication bias was analyzed by funnel plot., Results: A total of 28 RCTs were enrolled and 4 420 patients were included (2 207 in intervention group and 2 213 in control group). Meta-analysis showed that there was no significant difference in mortality between intervention group and control group [odds ratio (OR) = 0.90, 95% confidence interval (95%CI) = 0.75 to 1.08, P = 0.27]. The length of ICU stay [mean difference (MD) = -0.23, 95%CI = -0.29 to -0.16, P < 0.000 01] and the length of hospital stay (MD = -0.96, 95%CI = -1.21 to -0.70, P < 0.000 01) in intervention group were less than those in control group. Subgroup analysis showed that mortality of patients with sepsis and septic shock in intervention group was lower than that in control group (OR = 0.65, 95%CI = 0.43 to 0.99, P = 0.04). For patients undergoing cardiac surgery, the incidence of postoperative atrial fibrillation in intervention group was lower than that in control group (OR = 0.43, 95%CI = 0.34 to 0.54, P < 0.000 01). It was shown by funnel plot that there was less publication bias among studies., Conclusions: Vitamin C does not reduce mortality in critically ill patients, but it can reduce the length of ICU stay and hospital stay. In addition, vitamin C can reduce mortality of patients with sepsis and septic shock and reduce the incidence of atrial fibrillation post operation in patients undergoing cardiac surgery.

Objective: To systematically evaluate the effect of high-flow nasal cannula oxygen (HFNC) on improving the atelectasis and respiratory function in adults after cardiac surgeries., Methods: All randomized controlled trials (RCTs) about HFNC therapy for adults after cardiac surgeries published from January 2000 to March 2018 were searched through CNKI, CBM, VIP, Wanfang, PubMed, Springer Link, Embase, Web of Science, Cochrane Library. The references from relevant articles were searched. The experimental group was treated with HFNC while the control group treated with conventional oxygen therapy (COT). The outcome measurements included radiological atelectasis score (RAS), endotracheal reintubation rate and the length of intensive care unit (ICU) stay. Two researchers were responsible for literature screening, data extraction and quality evaluation respectively. Meta-analysis was performed with RevMan 5.2 software. Funnel plot was used to analyze the publication bias., Results: A total of 4 RCTs were enrolled and 643 patients were included (325 in experimental group and 318 in control group). Meta-analysis showed that the tracheal reintubation rate in experimental group was lower than that in control group [odds ratio (OR) = 0.26, 95% confidence interval (95%CI) = 0.09-0.74, P = 0.01], but there was no significant difference in RAS [mean difference (MD) = -0.15, 95%CI = -0.50-0.21, P = 0.41] and the length of ICU stay (MD = 0.09, 95%CI = -0.09-0.26, P = 0.33) between experimental group and control group. Sensitivity analysis was performed in two trials with low risk of bias, which demonstrated that there was no significant difference in RAS between the two groups (MD = 0.06, 95%CI = -0.26-0.37, P = 0.73). It was shown by the funnel analysis that there was bias in the study of the length of ICU stay in the literature, while the bias of RAS and tracheal reintubation rate was low., Conclusions: Compared with COT, HFNC could reduce the rate of tracheal reintubation in adults after cardiac surgeries, but no difference was found in improving atelectasis or reducing the length of ICU stay.

Olive flounder (Paralichthys olivaceus) is an economically important cultured marine fish in China, Korea, and Japan. Controlling and managing the breeding of olive flounder in captivity is an imperative step toward obtaining a sustainable supply of this fish in aquaculture production systems. Therefore, investigation on the molecular regulatory mechanism of gonadal development and gametogenesis in this species is of great significance in aquaculture. Furthermore, identification of the expression profile of numerous sex-related genes is the first step to primarily understand such molecular regulatory mechanism. Six female and six male gonads obtained from 2-year-old olive flounders were sequenced using Illumina, which produced 6.68 and 6.24 GB data for ovary and testis, respectively. The reads were mapped to the draft genome of olive flounder, and then the reads per kilobase per million (FPKM) for each gene were calculated. The female-/male-biased expressed genes were investigated based on the FPKM values. Overall, 3086 female-biased and 5048 male-biased genes were screened out. GO enrichment analysis showed that the GO terms "male meiosis," "gamete generation," "fertilization," "spermatogenesis," and "germ plasma" were enriched in male-biased genes. In addition, the GO terms "cell morphogenesis involved in differentiation," "embryonic morphogenesis," "plasma membrane," "steroid hormone receptor activity," and "aromatase activity" were enriched in female-biased genes. Moreover, 373,369 single nucleotide polymorphisms and 32,993 indels were identified in the transcriptome. This work is the largest collection of gonad transcriptome data for olive flounder and provides an extensive resource for future gonadal development and gametogenesis molecular biology studies in this species.

Sox2 has essential roles in early embryogenesis and the development of the central nervous system. Sox2 is also necessary in maintaining the identity of progenitor cells. In our study, a 1.8-kb fragment of the 5' flanking region of Paralichthys olivaceus Sox2 (Po-Sox2) gene was cloned and functionally characterized. The activity and specificity of Po-Sox2 promoter were analyzed by comparing various deletion mutants for their ability to direct luciferase and GFP expression in flounder brain cell line. Results indicated that the basal promoter is located between -978 and -442 bp, and the region from -1370 to -978 bp enhances the promoter activity. The regulatory elements in the -1370 to -442 bp fragment were further investigated. Many binding sites of transcription factors closely related to neurogenesis and stem cell properties were found in this region. Mutational analysis indicated that Nanog, Pax6, p53, and POU binding sites play functional roles in the transcription of Po-Sox2 gene, whereas NF-Y binding sites did not affect this gene. In vivo studies using transient transgenic zebrafish embryos showed that the Po-Sox2 promoter region can drive GFP expression in brain, yolk syncytial layer, and notochord. Our results provide valuable information in understanding the molecular regulatory mechanisms of Po-Sox2 gene during neurogenesis and embryonic development.

Following the two rounds of whole-genome duplication (WGD) during deuterosome evolution, a third genome duplication occurred in the ray-fined fish lineage and is considered to be responsible for the teleost-specific lineage diversification and regulation mechanisms. As a receptor-regulated SMAD (R-SMAD), the function of SMAD3 was widely studied in mammals. However, limited information of its role or putative paralogs is available in ray-finned fishes. In this study, two SMAD3 paralogs were first identified in the transcriptome and genome of Japanese flounder ( Paralichthys olivaceus ). We also explored SMAD3 duplication in other selected species. Following identification, genomic structure, phylogenetic reconstruction, and synteny analyses performed by MrBayes and online bioinformatic tools confirmed that smad3a/3b most likely originated from the teleost-specific WGD. Additionally, selection pressure analysis and expression pattern of the two genes performed by PAML and quantitative real-time PCR (qRT-PCR) revealed evidence of subfunctionalization of the two SMAD3 paralogs in teleost. Our results indicate that two SMAD3 genes originate from teleost-specific WGD, remain transcriptionally active, and may have likely undergone subfunctionalization. This study provides novel insights to the evolution fates of smad3a/3b and draws attentions to future function analysis of SMAD3 gene family., Competing Interests: The authors declare that they have no competing interests.

Objective: To investigate the diagnostic value of TruScreen (TS) as a novel option for cervical lesions screening., Methods: A total of 218 cases were selected between 2013 December and 2014 November from outpatients of the Department of Gynecology, PLA General Hospital. Using histopathology as the gold standard for diagnosis, the diagnostic efficacy of TS was compared with that of high-risk human papillomavirus (Hr-HPV) test., Results: (1) In 218 patients, there were 193 Hr-HPV positive cases accounting for 88.5%, 94 TS positive cases accounting for 43.1%. According to the histopathology, there were 112 cases of cervical intraepithelial neoplasia (CIN)1+, 71 cases of CIN2+, accounting for 51.4% and 32.6%, respectively. (2) The sensitivity of Hr-HPV was 98.2% and specificity was 21.7%. The sensitivity of TS was 51.8% and specificity 66.0%. (3) Sensitivity of TS for triaging cases of atypical squamous cells of undetermined signification (ASC-US) was 43.9%, specificity 65.7%, diagnostic accordance rate 53.9%. Those indexes for triaging cases of equivalent or more than atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion (ASC-H) were 56.3%, 66.2% and 61.3%. With regard to Hr-HPV test, those indexes for triaging cases of equivalent or more than ASC-H were 97.2%, 32.4% and 64.8%., Conclusions: Hr-HPV test shows higher sensitivity while TruScreen shows higher specificity for cervical lesions screening. More studies with larger sample size are still needed to evaluate which method is a better triage tool.

Following the two rounds of whole-genome duplication that occurred during deuterostome evolution, a third genome duplication event occurred in the stem lineage of ray-finned fishes. This teleost-specific genome duplication is thought to be responsible for the biological diversification of ray-finned fishes. DEAD-box polypeptide 3 (DDX3) belongs to the DEAD-box RNA helicase family. Although their functions in humans have been well studied, limited information is available regarding their function in teleosts. In this study, two teleost Ddx3 genes were first identified in the transcriptome of Japanese flounder (Paralichthys olivaceus). We confirmed that the two genes originated from teleost-specific genome duplication through synteny and phylogenetic analysis. Additionally, comparative analysis of genome structure, molecular evolution rate, and expression pattern of the two genes in Japanese flounder revealed evidence of subfunctionalization of the duplicated Ddx3 genes in teleosts. Thus, the results of this study reveal novel insights into the evolution of the teleost Ddx3 genes and constitute important groundwork for further research on this gene family., (Copyright © 2015 Wang et al.)

Background and Objective: It has been proven that increase of the awareness level of lung cancer prevention and control could enhance participation of lung cancer screening of lung cancer high risk group. The aim of this study is to investigate the awareness level of lung cancer prevention and control and the effect of individual characteristics on lung cancer awareness, and to provide evidence for comprehensive lung cancer prevention in high risk areas of lung cancer., Methods: Staffs of Tianjin Dagang Oil Field who participate low dose CT (LDCT) lung cancer screening by cluster sampling or according to voluntary principle were surveyed, data of lung cancer awareness were collected by questionnaire., Results: A total of 1,633 valid questionnaires were collected. The average age of respondents was 60.08±6.58. Most participants were males (82.2%) while female only accounted for 17.8%. The proportions of awareness about lung cancer in China, risk factors, screening methods and the knowledge of health examination were 64.5%, 77.1%, 43.7%, 49.6% respectively. Result of multiple logistic regression analysis showed that education level, smoking (pack-year), age, prior tuberculosis were the influencing factors of lung cancer awareness with adjusted Ors for education and age level as of 0.567 (95%CI: 0.439-0.733) and 1.373 (95%CI: 1.084-1.739) respectively. 80.3% of the participants can accept health examination once a year, while the ability to pay the medical expenses was not high. The influencing factors of health examination willingness were gender, age, income, the knowledge of lung cancer., Conclusions: Education level and smoking affect the awareness of lung cancer prevention and control, health education for lung cancer should be conducted especially in population with low education level. Comprehensive lung cancer control in high risk areas should combined lung cancer screening, tobacco control and health education.

Background and Objective: BAG genes (Bcl-2-associated athanogene) belong to a recently discovered multifunctional anti-apoptosis gene family that regulate various physiological processes which include apoptosis, tumorigenesis, neural differentiation, stress response and cell cycle and so on. The expression status of BAG family genes are related to certain tumor incidence and prognosis. The aim of this study is to explore the association of the BAG family gene expression status with the susceptibility of lung adenocarcinoma., Methods: The gene expression data of BAG family genes from 29 cases of lung adenocarcinoma tissues and matched pericancerous lung tissues were generated by microarray chips. Cox regression was used to analyze the association between the expression of BAG family genes and the susceptibility of lung adenocarcinoma and the results were verified by GEO database., Results: The expression levels of BAG-1, BAG-2, BAG-5 in cancer tissues were significantly downregulated compared with matched pericancerous lung tissues and were protective factors of lung adenocarcinoma (P < 0.05, OR < 1); while the expression level of BAG-4 in cancer tissues were remarkably upregulated compared with the matched pericancerous lung tissues and was risk factor of lung adenocarcinoma (P < 0.05, OR > 1)., Conclusions: BAG-1, BAG-2, BAG-5 might be the potential protective factors while BAG-4 is possible risk factor of lung adenocarcinoma.