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AIM: To compare the clinical curative effects of meibomian gland dysfunction(MGD)caused by evaporative dry eye by utilizing levofloxacin eye gel and Tobramycin and Dexamethasone Ophthalmic Ointment, respectively.METHODS: The 180 cases(360 eyes)with dry eye(evaporative type)caused by confirmed meibomian gland dysfunction were randomly divided into two groups(i.e. A and B): In group A, 90 individuals with 180 eyes were treated with levofloxacin eye gel+ sodium hyaluronate eye solution; The other 90 cases in group B took Tobramycin and Dexamethasone Ophthalmic Ointment+ sodium hyaluronate eye drops for curing the MGD. In addition, the same comprehensive therapy were used to the MGD patients in groups A and B, after surface anesthesia on binoculus, secretion, obstructing meibomian gland secretions, were discharged by utilizing cotton stick to extrusion mass weekly and four times consecutive treatments were regard as a course of treatment. To remove residual the thin oil soften lipid in meibomian gland, heat can be applied to the eyelids with hot water(around 45℃)on towel for 15min and do that three times a day. After each hot compress, we use levofloxacin eye gel to the patients in group A by dropping into the conjunctival sac and apply to the root of the eyelid lashes. The group B of 90 patients were applied Tobramycin and Dexamethasone Ophthalmic Ointment to the root of the eyelid lashes. All patients were dripped odium eye drops eye into their eyes four times a day. RESULTS: After treatment(Z= -0.64, P=0.524), there were no significant differences in clinical symptoms(Z= -1.37, P=0.171), secretion characteristics(Z= -1.06, P=0.288), tear film rupture time and tear secretion time between groups A and B(P>0.05). Corneal fluorescence staining score: group A(cured 83.3%, improved 11.1%, ineffective 5.6%)and group B(cured 55.6%, improved 27.8%, ineffective 16.7%). The therapeutic effect of group A was better than that of group B, with statistical significance(Z= -4.02, PCONCLUSION: Physical therapy for meibomian gland dysfunction caused by evaporative dry eye is given priority, and medication is treated as adjunctive therapy. Generally, the patients can achieve totally anti-inflammatory, antibacterial, safe and stable, without side effects by using levofloxacin eye gel. However, the patients with worst condition and lingering illness should cured by Tobramycin and Dexamethasone Ophthalmic Ointment. In addition, statistical significant difference is not found between the two drugs on curative effects.

Abstract The use of immunotherapy has achieved great advances in the treatment of cancer. Macrophages play a pivotal role in the immune defense system, serving both as phagocytes (removal of pathogens and cancer cells) and as antigen‐presenting cells (activation of T cells). However, research regarding tumor immunotherapy is mainly focused on the adaptive immune system. The usefulness of innate immune cells (eg, macrophages) in the treatment of cancer has not been extensively investigated. Recent advances in synthetic biology and the increasing understanding of the cluster of differentiation 47/signal regulatory protein alpha (CD47/SIRPɑ) axis may provide new opportunities for the clinical application of engineered macrophages. The CD47/SIRPɑ axis is a major known pathway, repressing phagocytosis and activation of macrophages. In this article, we summarize the currently available evidence regarding the CD47/SIRPɑ axis, and immunotherapies based on blockage. In addition, we propose cell therapy strategies based on macrophage engineering.

Objective To investigate the functional connectivity changes of benign epilepsy of childhood with centrotemporal spikes (BECTS) by using degree centrality (DC) analysis. Methods Choose 27 children diagnosed as BECTS from January 2010 to December 2017 as object of observation and 27 age/ sex⁃matched normal children as control. All subjects were performed fMRI in resting state. DC analysis was used to characterize the connectome patterns of whole⁃brain functional networks. Pearson correlation analysis was performed to evaluate the relationship between DC values of brain regions that differed between BECTS and normal children and the behavioral factors, including Raven's Standard Progressive Matrices (RSPM) intelligence quotient (IQ) and Integrated Visual and Auditory Continuous Performance Test (IVA⁃CPT). Results Compared with normal children, BECTS children exhibited higher DC valuesinthe right superior frontal gyrus and right posterior cingulate gyrus, and lower DC values in the bilateral cerebellar posterior lobes, subcortical mass in bilateral corpus striatum and/or thalami and right superior frontal gyrus. The higher DC value in the right posterior cingulate gyrus showed significant negative correlations with RSPM IQ score(r=⁃0.597, P=0.001) and full scale attention quotient score(r=⁃0.400, P=0.039). The RSPM IQ score showed significant positive correlations with the lower DC values in left cerebellar posterior lobe (r=0.442, P =0.021) and right corpus striatum/thalamus (r=0.452, P =0.018). Conclusions BECTS is associated with functional abnormalities in thalamic⁃cortical networks. DC analysis could provide a brand⁃new method for exploring the neurobiological mechanisms of epileptic discharges in BECTS.

Laccase is a multi‐copper oxidase which oxidizes substrate at the type 1 copper site, simultaneously coupling the reduction of dioxygen to water at the trinuclear copper center. In this study, we used site‐directed mutagenesis to study the effect of axial bonds between the metal and amino acid residue side chains in lacTT. Our kinetic and spectral data showed that the replacement of the axial residue with non‐coordinating residues resulted in higher efficiency (kcat/Km) and a lower Cu2+ population at the type 1 copper site, while substitution with strongly coordinating residues resulted in lower efficiency and a higher Cu2+ population, as compared with the wild‐type. The redox potentials of mutants with hydrophobic axial residues (Ala and Phe) were higher than that of the wild‐type. In conclusion, these insights into the catalytic mechanism of laccase may be of use in protein engineering to fine‐tune its enzymatic properties for industrial application.

AIM: To investigate microvascular changes in eyes with central retinal vein occlusion (CRVO) complicated by macular edema before and after intravitreal conbercept injection and evaluate correlations between these changes and best-corrected visual acuity (BCVA) and retinal thickness. METHODS: Twenty-eight eyes of 28 patients with macular edema caused by CRVO were included in this retrospective study. All patients received a single intravitreal conbercept injection to treat macular edema. BCVA and the results of optical coherence tomography angiography (OCTA) automatic measurements of the vessel density in the superficial (SCP) and deep retinal capillary plexus (DCP), the foveal avascular zone (FAZ) area, the FAZ perimeter (PERIM), the vessel density within a 300-μm wide ring surrounding the FAZ (FD-300), the acircularity index (AI), the choriocapillaris flow area, and retinal thickness were recorded before and at one month after treatment and compared with the results observed in age- and sex- matched healthy subjects. RESULTS: The vessel density in the SCP and DCP, the FD-300, and the flow area of the choriocapillaris were all significantly lower in CRVO eyes than in healthy eyes, while the AI and retinal thickness were significantly higher (all P

Abstract Genetic testing for germline mutations in BRCA1/2 of patients with breast cancer (BC) is part of routine patient care. However, BRCA1/2 mutations account only for a fraction of familial BC. A custom panel of 22 gene sequencing was performed on each patient. Among the 481 female patients, 135 patients were detected to carry pathogenic (P)/likely pathogenic (LP) mutations (28.1%), which corresponded to 12 different cancer predisposition genes [14.6% (70/481) on BRCA1 gene, 5.0% (24/481) on BRCA2 gene, 8.5% (41/481) on non‐BRCA1/2 genes]. Moreover, 24.7% (119/481) of patients had mutation of unknown significance (VUS) in these genes. The most common (8/481) pathogenic mutation is BRCA1 c.5470_5477del, while BRIP1 2392 C > T of patients was detected. All the mutations detected were mainly seen in the homologous recombinant repair pathway. Compared to BRCA2 mutation, BRCA1 mutation is higher in younger female patients (P

Abstract Background Breast cancer angiogenesis is key for metastasis and predicts a poor prognosis. Angiotensin-converting enzyme 2 (ACE2), as a member of the renin-angiotensin system (RAS), was reported to restrain the progression of hepatocellular carcinoma (HCC) and non-small cell lung cancer (NSCLC) through inhibiting angiogenesis. However, the relationship between ACE2 and breast cancer angiogenesis remains unclear. Methods The prognosis and relative gene selection were analysed using the GEPIA, GEO, TCGA and STRING databases. ACE2 expression in breast cancer tissue was estimated by reverse transcription-quantitative polymerase chain reaction (qPCR). Breast cancer cell migration, proliferation and angiogenesis were assessed by Transwell migration, proliferation, tube formation, and wound healing assays. The expression of vascular endothelial growth factor A (VEGFa) was detected by qPCR and Western blotting. The phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2), mitogen-activated protein kinase 1/2 (MEK1/2), and extracellular signal-regulated protein kinase 1/2 (ERK1/2) was examined by Western blotting. Breast cancer metastasis and angiogenesis in vivo were measured using a zebrafish model. Results ACE2 was downregulated in breast cancer patients. Patients with higher ACE2 expression had longer relapse-free survival (RFS). In vitro, ACE2 inhibited breast cancer migration. Meanwhile, ACE2 in breast cancer cells inhibited human umbilical vascular endothelial cell (HUVEC) proliferation, tube formation and migration. In the zebrafish model, ACE2 inhibited breast cancer cell metastasis, as demonstrated by analyses of the number of disseminated foci and the metastatic distance. Neo-angiogenesis was also decreased by ACE2. ACE2 downregulated the expression of VEGFa in breast cancer cells. Furthermore, ACE2 in breast cancer cells inactivated the phosphorylation of VEGFR2, MEK1/2, and ERK1/2 in HUVECs. Conclusions Our findings suggest that ACE2, as a potential resister to breast cancer, might inhibit breast cancer angiogenesis through the VEGFa/VEGFR2/ERK pathway. Trial registration Retrospectively registered.

Abstract Background Noninvasive prenatal screening (NIPS) based on cell-free fetal DNA (cffDNA) has rapidly been applied into clinic. However, the reliability of this method largely depends on the concentration of cffDNA in the maternal plasma. The chance of test failure results or false negative results would increase when cffDNA fraction is low. In this study, we set out to develop a method to enrich the cffDNA for NIPS based on the size difference between cell-free DNA (cfDNA) of fetal origin and maternal origin, and to evaluate whether the new NIPS method can improve the test quality. Methods We utilized 10,000 previous NIPS data to optimize a size-selection strategy for enrichment. Then, we retrospectively performed our new NIPS method with cffDNA enrichment on the 1415 NIPS samples, including 1404 routine cases and 11 false negative cases, and compared the results to the original NIPS results. Results The 10,000 NIPS data revealed the fetal fraction in short cfDNA fragments (

Abstract Background Translational regulation played an important role in the correct folding of heterologous proteins to form bioactive conformations during biogenesis. Translational pausing coordinates protein translation and co-translational folding. Decelerating translation elongation speed has been shown to improve the soluble protein yield when expressing heterologous proteins in industrial expression hosts. However, rational redesign of translational pausing via synonymous mutations may not be feasible in many cases. Our goal was to develop a general and convenient strategy to improve heterologous protein synthesis in Pichia pastoris without mutating the expressed genes. Results Here, a large-scale deletion library of ribosomal protein (RP) genes was constructed for heterologous protein expression in Pichia pastoris, and 59% (16/27) RP deletants have significantly increased heterologous protein yield. This is due to the delay of 60S subunit assembly by deleting non-essential ribosomal protein genes or 60S subunit processing factors, thus globally decreased the translation elongation speed and improved the co-translational folding, without perturbing the relative transcription level and translation initiation. Conclusion Global decrease in the translation elongation speed by RP deletion enhanced co-translational folding efficiency of nascent chains and decreased protein aggregates to improve heterologous protein yield. A potential expression platform for efficient pharmaceutical proteins and industrial enzymes production was provided without synonymous mutation.

Objective: To investigate the effects of gestational diabetes mellitus (GDM) on time to delivery and perinatal outcomes in full-term pregnancies underwent dinoprostone-induced labor. Methods: GDM patients that underwent labor induction with dinoprostone vaginal inserts were retrospectively recruited. Full-term pregnancies with normal glucose tolerance (NGT) that underwent labor induction at the same period were recruited as control. Time to delivery and perinatal outcomes were compared between the two groups. Results: A total of 1555 pregnancies with 226 GDM and 1329 NGT were recruited. GDM pregnancies had older ages, lower gestational age, higher body mass index (BMI) and abortion history, and more multigravida than NGT pregnancies (P< 0.05). Univariate analysis showed no significant difference in time to delivery and delivery rates between the two groups. However, after adjusted in a multivariate analysis model, the delivery rates of GDM women delivered within 12, 24, 36 or 48 h and those vaginally delivered within 12 or 36 h were significantly lower than those in the NGT group (P< 0.05). Perinatal outcomes were clinically similar between the two groups. Conclusion: GDM did not affect the time to delivery, cesarean delivery and other perinatal outcomes in Chinese women underwent dinoprostone-induced labor. However, it may be associated with the lower rates of delivery within different time intervals.

Distributed cooperative co-evolution (DCC) is an effective way to solve large-scale optimization problems. The cooperative co-evolution methodology can reduce the optimizing complexity by dividing a large-scale problem into small subcomponents, and distributed computation can accelerate the optimizing speed. However, existing DCC algorithms often encounter deficiency in overlapping problems that cannot be ideally divided due to unavoidable overlaps between subcomponents. To address this issue, this paper proposes an algorithm called distributed cooperative co-evolutionary covariance matrix adaptation evolution strategy (DCCMAES) to solve large-scale overlapping optimization problems. First, a new grouping scheme is introduced, where a variable, if needed, can be assigned into multiple subcomponents. Second, to address the conflicts caused by optimizing multiple copies of shared variables, a random orthogonal experiment method is proposed to generate global solutions. Moreover, three cooperation schemes are proposed to coordinate the optimizers in DCCMAES during evolution. The experiments on benchmark functions and a real-world application show that DCCMAES is promising for global optimization of large-scale overlapping problems.

Demand side management is an efficient way of reducing the grid fluctuation and enhancing the net profit in the smart grid. As the increasing complexity of the real-world applications, e.g., NP-hard and non-polynomial problems, a distributed coevolution algorithm is proposed to cope with this challenge. In the proposed algorithm, the management system is modeled as a hierarchical structure. The user side locates at the lowest level and upper levels represent the system operators. The system operators manage users' energy consumption behavior through the real-time pricing strategy. The genetic algorithm and particle swarm optimization are modified to play the role of the smart scheduler and the smart pricing generator. The end users conduct appliance commitment using the smart scheduler which considers the users' comfort level and the electricity cost. The decision of the lower level depends directly on the real-time pricing strategy released by its adjacent upper level. The real-time pricing is developed by the smart pricing generator which considers the net profit and the grid fluctuation. Three types of experiments are conducted on a distributed platform to investigate and ascertain the performance of the proposed algorithm. For the operator side, the experimental results indicate that the proposed algorithm improves the grid fluctuation, and enhances the net profit. As for the user side, it improves the comfort level and achieves budget saving.

Uncovering community structure is an important technique for studying complex networks. While a large bulk of algorithms have been proposed for community detection in recent years, most of them were designed for undirected networks. Considering many real-world networks are by nature directed, it is necessary to develop community detection methods that can handle directed networks. In this work, we formulates a multi-objective framework for community detection in directed networks and proposes a multi-objective evolutionary algorithm for finding efficient solutions under this framework. Specifically, based on the theory that an efficient partition of directed networks should have larger network information flow within the community than that between different communities, we first designed two conflicting objective functions based on PageRank random walk, one of which is to maximize within-community transition probability, and the other is to minimize between-community transition probability. By optimizing these two objectives simultaneously, we modelled the problem of community detection as a multi-objective optimization problem, and then developed a novel multi-objective evolutionary algorithm to solve it. Particularly, to guarantee the capability of searching the optimal solution, our proposed method designed/adopted the directed-network-specific population initialization method and evolutionary operator by introducing label propagation algorithm into multi-objective genetic algorithm. Comparison with other four art-of-the-state algorithms, our method showed the competitive performance on both synthetic and real-world networks. Moreover, attributing to the multi-objective framework, the proposed method could generate multiple optimal network partitions in a single run, which provides a hierarchical description of community structure of the network.

Impact of high-speed jet (HSJ) treatment on morphology, crystalline structure, retrogradation, thermal and rheological properties of Arenga pinnata (Wurmb.) Merr starch (APS) was investigated. Decreasing with the number of HSJ treatment passes were the turbidity values of pastes (long-term retrogradation), the gelatinization enthalpy, and the crystallinity. Scanning electron microscopy showed APS granule structure was degraded into tiny round particle and thin sheet structures. X-ray diffraction illustrated that the C-type X-ray pattern was changed toward a weak A-type X-ray pattern. With further increases in the number of passes, the weak A-type pattern was changed to amorphous pattern. Dynamic oscillation analysis revealed that HSJ treatment significantly altered the viscoelasticity of APS. During a dynamic frequency sweep, G′ values and G″ at 10 Hz decreased from 589.9 Pa (HSJ0) to 194.9 Pa (HSJ8) and from 101.6 Pa (HSJ0) to 24.5 Pa (HSJ8), respectively, implying that HSJ treatment had a great influence on the elasticity of the APS gel. The results indicated that degradation of APS granular and crystalline structure was mainly involved and HSJ treatment was a potential physical modification technique.

Abstract Background To examine the influence of HOXD10 on the metabolism and growth of colon carcinoma cells by suppressing the RHOC/AKT/MAPK pathway. Methods Thirty-seven paired colon cancer and its adjacent samples from The Cancer Genome Atlas (TCGA) were analyzed. Chip Analysis Methylation Pipeline (ChAMP) analysis was employed for differential methylated points (DMPs) and the differential methylation regions (DMRs) screening. The HOXD10 mRNA expression and DNA methylation levels were detected by RT-PCR. The Cell proliferation, migration, invasion and apoptosis were respectively measured by MTT assay, transwell assay, wound healing assay and flow cytometry assay in carcinoma cell lines after treated with 5-aza-2′-deoxycytidine (5-Aza-dC) or transfected with HOXD10-expressing plasmid. The expression of HOXD10 and RHOC was revealed by immunohistochemistry in disparate differentiation colon carcinoma tissues, and the dephosphorylation of AKT and MAPK pathways were detected by RT-PCR and western blot. Results The bioinformatics analysis demonstrated that HOXD10 was hypermethylated and low-expressed in colorectal cancer tissues. The detection of RT-PCR indicated the similar results in colorectal cancer cell lines and tissues. The induction of demethylation was recovered by treatment with 5-Aza-dC and the HOXD10 in colorectal cancer cell lines was re-expressed by transfection with a HOXD10 expression vector. The demethylation or overexpression of HOXD10 suppressed proliferation, migration, invasion and promoted apoptosis in colorectal cancer cells. HXOD10 suppressed the tumor growth and detected an opposite trend of protein RHOC. AKT and MAPK pathways were notably inactivated after the dephosphorylation due to the overexpression of HOXD10. Conclusions HOXD10 was suppressed in colon adenocarcinoma cells, which down-regulated RHOC/AKT/MAPK pathway to enhance colon cancer cells apoptosis and constrain the proliferation, migration and invasion.

The estimation of non-stationary random medium parameters of petrophysical parameters is the key to the application of random medium theory in fine seismic exploration. We proposed a method for estimating non-stationary random medium parameters of petrophysical parameters using seismic data. Based on the linear petrophysical model, the relationship between seismic data and porosity, clay volume, and water saturation in the random medium was described, and the principle and method of estimating the autocorrelation parameters of the petrophysical parameter random medium were introduced in this study. Subsequently, the specific steps of applying the power spectrum method, for parameter estimation in non-stationary random media with petrophysical parameters, were explained. The feasibility and correctness of the method were verified through the estimation test of the two-dimensional theoretical model. Eventually, the estimation test of non-stationary random medium parameters of petrophysical parameters was carried out by field seismic data, and the results indicated that the non-stationary random medium parameters can better portray the information of subsurface medium petrophysical parameters. The method can provide a reference for the construction of a priori information on petrophysical parameters, and it can also provide a theoretical basis for the in-depth application of random medium theory to practical data.

Exposure to titanium dioxide nanoparticles (TiO2 NPs) and bisphenol A (BPA) is ubiquitous, especially through dietary and other environmental pathways. In the present study, adult C57BL/6J mice were exposed to TiO2 NPs (100 mg/kg), BPA (0, 5, and 50 mg/kg), or their binary mixtures for 13 weeks. The 16S rDNA amplification sequence analysis revealed that co-exposure to TiO2 NPs and BPA altered the intestinal microbiota; however, this alteration was mainly caused by TiO2 NPs. Faecal metabolomics analysis revealed that 28 metabolites and 3 metabolic pathways were altered in the co-exposed group. This study is the first to reveal the combined effects of TiO2 NPs and BPA on the mammalian gut microbial community and metabolism dynamics, which is of great value to human health. The coexistence of TiO2 NPs and BPA in the gut poses a potential health risk due to their interaction with the gut microbiota.

In the original publication [...]

Bisphenol A (BPA) and heavy metals are widespread contaminants in the environment. However, the combined toxicities of these contaminants are still unknown. In this study, the bioluminescent bacteria Vibrio qinghaiensis Q67 was used to detect the single and combined toxicities of BPA and heavy metals, then the joint effects of these contaminants were evaluated. The results show that chronic toxicities of chromium (Cr), cadmium (Cd), lead (Pb), arsenic (As), mercury (Hg), nickel (Ni), and BPA were time–dependent; in fact, the acute toxicities of these contaminants were stronger than the chronic toxicities. Furthermore, the combined toxicities of BPA and heavy metals displayed BPA + Hg > BPA + Cr > BPA + As > BPA + Ni > BPA + Pb > BPA + Cd in the acute test and BPA + Hg > BPA + Cd > BPA + As > BPA + Cd in the chronic test, which suggested that the combined toxicity of BPA and Hg was stronger than that of other mixtures in acute as well as chronic tests. Additionally, both CA and IA models underestimated the toxicities of mixtures at low concentrations but overestimated them at high concentrations, which indicates that CA and IA models were not suitable to predict the toxicities of mixtures of BPA and heavy metals. Moreover, the joint effects of BPA and heavy metals mainly showed antagonism and additive in the context of acute exposure but synergism and additive in the context of chronic exposure. Indeed, the difference in the joint effects on acute and chronic exposure can be explained by the possibility that mixtures inhibited cell growth and luminescence in chronic cultivation. The chronic toxicity of the mixture should be considered if the mixture results in the inhibition of the growth of cells.

Abstract Oral cancer remains a deadly disease worldwide. Lymph node metastasis and invasion is one of the causes of death from oral cancer. Elucidating the mechanism of oral cancer lymph node metastasis and identifying critical regulatory genes are important for the treatment of this disease. This study aimed to identify differentially expressed genes (gene signature) and pathways that contribute to oral cancer metastasis to lymph nodes. The GSE70604-associated study compared gene profiles in lymph nodes with metastasis of oral cancer to those of normal lymph nodes. The GSE2280-associated study compared gene profiles in primary tumor of oral cancer with lymph node metastasis to those in tumors without lymph node metastasis. There are 28 common differentially expressed genes (DEGs) showing consistent changes in both datasets in overlapping analysis. GO biological process and KEGG pathway analysis of these 28 DEGs identified the gene signature CCND1, JUN and SPP1, which are categorized as key regulatory genes involved in the focal adhesion pathway. Silencing expression of CCND1, JUN and SPP1 in the human oral cancer cell line OECM-1 confirmed that those genes play essential roles in oral cancer cell invasion. Analysis of clinical samples of oral cancer found a strong correlation of these genes with short survival, especially JUN expression associated with metastasis. Our study identified a unique gene signature – CCND1, JUN and SPP1 – which may be involved in oral cancer lymph node metastasis.

BackgroundLung squamous cell carcinoma (LUSC) is a major subtype of non-small cell lung cancer. The tumor immune microenvironment (TIME) affects the anti-tumor immune response and the patient’s prognosis, although the TIME in LUSC patients is incompletely understood.MethodsWe retrospectively collected surgical specimens from patients with previously untreated primary LUSC. Histopathological examination was used to identify tumor regions and adjacent regions, and imaging mass cytometry was used to characterize the immune cells in those regions. The results were compared between regions and between patients.ResultsWe identified heterogeneity in the TIME on comparing different patients with LUSC, although the tumor region and adjacent region both exhibited an immune response to the tumor. The TIME typically included a large number of infiltrating and activated T-cells, especially CD8+ T-cells, which closely interacted with the tumor cells in the tumor region. There was limited infiltration of B-cells, NK cells, and NKT cells, while the major immune suppressor cells were CD33+ myeloid-derived cells. We also identified a novel population of CD3−CD4+ cells with high expression of Foxp3 and TNFα, which might modulate the tumor microenvironment and play a proinflammatory role in the TIME.ConclusionsThe TIME of LUSC appears to be immunogenic and heterogenous, with predominant infiltration of activated CD8+ T-cells. The interactions between the tumor cells and T-cells facilitate the anti-tumor activity. A novel subpopulation of CD3−CD4+ cells with high TNFα and Foxp3 expression may modulate the tumor microenvironment and play a proinflammatory role.

The guarantee of cell survival under hypoxic conditions and rapid vascularization is a key in tissue engineering strategies for treating bone defects. Our study aimed to establish the protective role of bone marrow mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) in hypoxic conditions and realize rapid vascularization in bone defects. Resveratrol (Res), a non-flavonoid polyphenolic compound, and angiopoietin-2 (ANG2), a vascular activating factor, were applied to enhance BMSC and HUVEC survival, osteogenesis, and angiogenesis. The morphology, autophagy, viability, apoptosis, cycle, and osteogenic differentiation of BMSCs treated with Res were analyzed. The results indicated that Res could improve BMSC survival and differentiation via the autophagy pathway under hypoxic conditions. In addition, Res maintained HUVEC growth and proliferation in a hypoxic and ANG2 double-adverse environment via the autophagy pathway. To simulate a relatively hypoxic environment, small-aperture PEGDA/TCS hydrogels containing Res and ANG2 were prepared. BMSCs were cultured in the PEGDA/TCS scaffold and transplanted into a large tibial defect. CD31 immunofluorescence showed that the density and size of new blood vessels in the bone defect were significantly enhanced by ANG2 and Res at 8 weeks after surgery. H&E, Masson, and immunohistochemical staining results indicated that ANG2 combined with Res could promote new bone formation in defects. All these results suggested that Res combined with ANG2 may be a novel strategy for the targeted therapy of hypoxic bone defects with tissue engineering scaffolds.

Background: Although dual blockade HER2-based neoadjuvant chemotherapy is associated with excellent outcomes for human epidermal growth factor receptor 2 (HER2)-positive breast cancer, pertuzumab is not available to all patients due to cost. The optimal neoadjuvant chemotherapy for HER2-positive breast cancer in the presence of a single HER2 blockade is unknown. This study aimed to compare the efficacy and safety of epirubicin/cyclophosphamide followed by docetaxel/trastuzumab (EC-TH) with docetaxel/carboplatin/trastuzumab (TCH) neoadjuvant setting for HER2-positive breast cancer under the single HER2 blockade. Methods: Patients with stage II-IIIC HER2-positive breast cancer were randomly assigned to either eight cycles of EC-TH every 3 weeks during all chemotherapy cycles, or six cycles of TCH every 3 weeks. The primary endpoint was pathological complete response (pCR) (defined as the absence of invasive tumor cells in breast and axilla, ypT0/is ypN0). Results: From May 2017 to November 2019, 140 patients were randomly assigned, and 135 patients were ultimately found evaluable for the primary endpoint. The pCR was recorded in 25 of 67 patients [37.3%; 95% confidence interval (CI), 25.8–50.0] in the EC-TH group and in 38 of 68 patients (55.9%, 95% CI, 43.3–67.9) in the TCH group ( p = 0.032). The most common adverse events (AEs) were neutropenia in 24 of 67 (35.8%) patients in the EC-TH group versus 27 of 68 (39.7%) in the TCH group ( p = 0.642), anemia in 33 of 67 (49.3%) patients in the EC-TH group versus 34 of 68 (50.0%) in the TCH group ( p = 0.931), and thrombocytopenia in five of 67 (7.5%) patients in the EC-TH group versus 17 of 68 (25.0%) in the TCH group ( p = 0.006). Conclusion: For patients receiving the single HER2 blockade trastuzumab for HER2-positive breast cancer, TCH regimen might be a preferred neoadjuvant therapy. Trial registration: This trial was registered with ClinicalTrials.gov identifier: NCT03140553) on 2 May 2017.

BackgroundNeural tube defects (NTDs) are among the most common and severe congenital defects in humans. Their genetic etiology is complex and remains poorly understood. The Mediator complex (MED) plays a vital role in neural tube development in animal models. However, no studies have yet examined the role of its human homolog in the etiology of NTDs.MethodsIn this study, 48 pairs of neural lesion site and umbilical cord tissues from NTD and 21 case-parent trios were involved in screening for NTD-related somatic and germline de novo variants. A series of functional cell assays were performed. We generated a Med12 p.Arg1784Cys knock-in mouse using CRISPR/Cas9 technology to validate the human findings.ResultsOne somatic variant, MED12 p.Arg1782Cys, was identified in the lesion site tissue from an NTD fetus. This variant was absent in any other normal tissue from different germ layers of the same case. In 21 case-parent trios, one de novo stop-gain variant, MED13L p.Arg1760∗, was identified. Cellular functional studies showed that MED12 p.Arg1782Cys decreased MED12 protein level and affected the regulation of MED12 on the canonical-WNT signaling pathway. The Med12 p.Arg1784Cys knock-in mouse exhibited exencephaly and spina bifida.ConclusionThese findings provide strong evidence that functional variants of MED genes are associated with the etiology of some NTDs. We demonstrated a potentially important role for somatic variants in the occurrence of NTDs. Our study is the first study in which an NTD-related variant identified in humans was validated in mice using CRISPR/Cas9 technology.

Electroencephalography (EEG)-based driving fatigue detection has gained increasing attention recently due to the non-invasive, low-cost, and potable nature of the EEG technology, but it is still challenging to extract informative features from noisy EEG signals for driving fatigue detection. Radial basis function (RBF) neural network has drawn lots of attention as a promising classifier due to its linear-in-the-parameters network structure, strong non-linear approximation ability, and desired generalization property. The RBF network performance heavily relies on network parameters such as the number of the hidden nodes, number of the center vectors, width, and output weights. However, global optimization methods that directly optimize all the network parameters often result in high evaluation cost and slow convergence. To enhance the accuracy and efficiency of EEG-based driving fatigue detection model, this study aims to develop a two-level learning hierarchy RBF network (RBF-TLLH) which allows for global optimization of the key network parameters. Experimental EEG data were collected, at both fatigue and alert states, from six healthy participants in a simulated driving environment. Principal component analysis was first utilized to extract features from EEG signals, and the proposed RBF-TLLH was then employed for driving status (fatigue vs. alert) classification. The results demonstrated that the proposed RBF-TLLH approach achieved a better classification performance (mean accuracy: 92.71%; area under the receiver operating curve: 0.9199) compared to other widely used artificial neural networks. Moreover, only three core parameters need to be determined using the training datasets in the proposed RBF-TLLH classifier, which increases its reliability and applicability. The findings demonstrate that the proposed RBF-TLLH approach can be used as a promising framework for reliable EEG-based driving fatigue detection.

Introduction Diabetic retinopathy (DR) is the main cause of adult visual impairment worldwide. Severe non-proliferative DR (sNPDR) is an important clinical intervention stage. Currently, panretinal photocoagulation (PRP) is the standard treatment for sNPDR. However, PRP alone cannot completely prevent NPDR progression. One explanation might be that PRP does not remove the detrimental vitreous that plays an important role in DR progression. Microinvasive pars plana vitrectomy (PPV) was shown to be a safe and effective method to treat late-stage proliferative DR (PDR) by completely removing the pathological vitreous. However, whether PPV is effective in controlling sNPDR remains unknown. In this trial, we aim to compare the effectiveness of microinvasive PPV with that of PRP for sNPDR progression control.Methods and analysis This single centre, parallel group, randomised controlled trial aims to evaluate the clinical efficacy of microinvasive PPV in preventing the progression of sNPDR compared with PRP. A total of 272 adults diagnosed with sNPDR will be randomised 1:1 to the microinvasive PPV and PRP groups. The primary outcome is the disease progression rate, calculated as the rate of sNPDR progressed to PDR from baseline to 12 months after treatment. The secondary outcomes include the change in best-corrected visual acuity, re-treatment rate, diabetic macular oedema occurrence, change in central retinal thickness, change in the visual field, cataract occurrence and change in the quality of life.Ethics and dissemination The Ethics Committee of Zhongshan Ophthalmic Center approved this study (2019KYPJ108). The results will be presented at scientific meetings and submitted for publication to peer-reviewed journals.Trial registration number NCT04103671.

Introduction: Uremic pruritus (UP) is one of the most bothersome symptoms among chronic kidney disease (CKD) patients. The pathophysiology of UP remains elusive, resulting in limited treatment options. The inability of standard medical treatments to provide effective relief has piqued interest in complementary and alternative medicine (CAM). Methodology: A systematic review of randomized controlled trials (RCTs) summarizing the efficacy and safety profile of CAM used for UP in CKD patients was performed. CAM interventions were classified using categories proposed by the National Center for Complementary and Integrative Health. The efficacy of each CAM was determined from changes in UP severity and all reported adverse effects were extracted. Results: Of 5242 articles screened, 34 RCTs were included, with 15 (44.1 %) studies having a sample size greater than 50. The studies considered 21 treatments including omega-3 fatty acid supplementation (n=5), acupuncture (n=5), topical capsaicin (n=4) and acupressure (n=3). Acupuncture, acupressure and topical capsaicin were shown to be effective in improving uremic pruritus. Interventions which include oral omega-3 fatty acid and zinc supplementation demonstrated mixed efficacy. Other therapies such as evening primrose oil, turmeric, vitamin B3, vitamin D and thermal therapy were not effective for treatment of UP. Common adverse effects reported with topical capsaicin included mild burning sensations (50.0–88.2 %) or erythema (6.7–22.7%) while that of acupuncture included soreness (7.5 %), bleeding (6.0–7.5%) and hematoma (1.9 %). Conclusions: Acupuncture, acupressure and topical capsaicin have the largest body of evidence for efficacy in the treatment of UP. Larger and higher quality RCTs are required to examine the efficacy and safety of promising CAM.

The identification of Chinese herbal medicines (CHMs) is directly related to their safety and efficacy. However, the limited number of professionals fails to meet the demand of the vast CHMs market. To make the CHMs identification more convenient and accurate, this study aimed at assessing the feasibility of the automated machine learning (AutoML) technology in CHMs image recognition. Instead of a handcrafted neural network experimental models built on AutoML platform were presented. A dataset of 31,460 images consisting of 315 categories of commonly-used CHMs was built for the model creation. Furthermore, the Huawei ModelArts model was compared with a model built on the Baidu EasyDL platform using the same dataset. Three professionals were also invited to recognize images of 315 categories of CHMs. During the model evaluation, high accuracies of 99.2% and 98.4% were achieved by ModelArts and EasyDL, respectively. In the held-out tests, the accuracies of ModelArts and EasyDL models were 91.2% and 91.85%, respectively. Both models performed very well individually and no statistically significant difference was found in model performance between them. However, the model-training time was only approximately 41 min on ModelArts platform but 118 min on EasyDL. The mean accuracy of the manual recognition for 315 CHMs was 97.46 ± 1.58%. Results revealed that AutoML technology is a fast and simple approach and has great practical potential in the field of CHMs image recognition. Since the Huawei ModelArts platform requires less training time, we recommend it as a priority.

Chitosan-based intelligent artificial systems have been of increasing interest for their biocompatibility, multifunctionality, biological activity, and low cost. Herein, we report the fabrication of supramolecular nanoparticles based on water-soluble chitosan (WCS) and 1,1′,1″,1″′-(ethene-1,1,2,2-tetrayl)tetrakis(benzene-4,1-diyl) tetrakis(azanediyl)tetraacetic acid (TPE-(N-COOH)4), which is capable of reversible transition between polyion complexes (PICs) and hydrogen bonding complexes (HBCs) with tunable aggregation-induced emission driven by pH value. The PIC micelles could be formed via electrostatic interaction between ammonium cations and carboxylate anions under mild alkaline conditions. The formation of the micelles dramatically blocks the nonradiative pathway and enhances the fluorescence of TPE moieties, and the maximum fluorescence intensity was achieved near the isoelectric point due to the restriction of intramolecular motion. In addition, the fluorescence intensity and size of the PIC micelles exhibited a temperature response in the range from 20 to 80°C. Upon adjusting the solution pH to 2, the PIC micelles were reconstructed into hydrogen-bonding complexes while the hydrogen bonding interaction between the protonated carboxyl groups of TPE-(N-COOH)4 and chitosan. Moreover, the size of the micelles underwent a remarkable decrease, whereas the fluorescence emission was further enhanced by ~6.25-fold. The pH actuated micellar transition from PIC to HBC with tunable fluorescence performance is fully reversible. This study provides novel multifunctional materials that are of great importance for their potential application in the fields of optoelectronic devices and chemical and biomedical sensors.

The microRNA-200 (miR-200) family has been reported to be vital for the inhibition of epithelial-to-mesenchymal transition (EMT) in tumor cells. The miR-200 family represents a complex multi-factorial regulatory network which has not been well described in breast cancer. This study aimed to clarify the underlying regulatory association between IL-8 and miR-200 family in the process of EMT in breast cancer cell. In estrogen-receptor (ER) positive breast cancer cell line MCF-7, IL-8 overexpression cells were performed by lentivirus transfection as endogenous regulation with additional exogenous IL-8 stimulation. Transient overexpressions of miR-200 family were performed after endogenous or exogenous IL-8 overexpression in MCF-7 cells. IL-8 knockdown cells were constructed via siRNA and shRNA transfection in triple negative breast cancer cell line MDA-MB-231. N-cadherin, vimentin and ZEB2 were down-regulated and E-cadherin was up-regulated in IL-8 knockdown group compared with control group. On the other hand, N-cadherin, vimentin and ZEB2 were up-regulated and E-cadherin was down-regulated in IL-8 overexpression group compared with control group. This indicated IL-8 promotes EMT in breast cancer cells. Transwell assay showed that IL-8 increased the migration and invasiveness of tumor cells. Furthermore, we performed transient overexpression of miR-200 family after endogenous or exogenous IL-8 overexpression in MCF-7 cells, which showed that the miR-200 family could inhibit EMT induced by IL-8. IL-8 promoted EMT via downregulation of miR-200 family expression in breast cancer cells and increases tumor cell migration and invasion.

Objective: This study aims to investigate perinatal depression in women who gave birth during the COVID-19 pandemic in Wuhan, and to evaluate the effect of the pandemic on perinatal depression prevalence.Methods: A cross-sectional investigation was conducted into women hospitalized for delivery in Hubei Maternity and Child Healthcare Hospital from December 31, 2019 to March 22, 2020, a period which encompasses the entire time frame of the COVID-19 pandemic in Wuhan. The Edinburgh Postnatal Depression Scale (EPDS) was adopted to evaluate perinatal depression status. A Chi-square test and logistic regression model were utilized for data analysis.Results: A total of 2,883 participants were included, 33.71% of whom were found to suffer from depressive symptoms. In detail, 27.02%, 5.24%, and 1.46% were designated as having mild, moderate, and severe depressive symptoms, respectively. The perinatal depression prevalence increased as the COVID-19 pandemic worsened. Compared to the period from December 31, 2019 to January 12, 2020, perinatal depression risk significantly decreased within the 3 weeks of March 2–22, 2020 (1st week: OR = 0.39, 95% CI: 0.20, 0.78; 2nd week: OR = 0.35, 95% CI: 0.17, 0.73; and 3rd week: OR = 0.48, 95% CI: 0.25, 0.94); and the postnatal depression risk significantly rose within the four weeks of January 27-February 23, 2020 (1st week: OR = 1.78, 95% CI: 1.18, 2.68; 2nd week: OR = 2.03, 95% CI: 1.35, 3.04; 3rd week: OR = 1.48, 95% CI: 1.02, 2.14; and 4th week: OR = 1.73, 95% CI: 1.20, 2.48).Conclusion: The dynamic change of perinatal depression was associated with the progression of the COVID-19 pandemic among new mothers who were exposed to the pandemic. An elevated risk of postnatal depression was also observed during the COVID-19 pandemic.

Growth hormone (GH) participates in many physiological processes through the activation of its membrane-anchored receptor (GHR) in vertebrates. Because of its pivotal role in regulating growth and development, a large number of studies examining GH-GHR have been conducted in mammals, birds and fish. However, whether a functional GH-GHR pair exists in the Chinese soft-shelled turtle (st-) has not been previously studied. Here, the full-length cDNA encoding GH was cloned from Chinese soft-shelled turtle (Trionyx sinensis) pituitary. The results revealed that it consisted of 784 bp and encoded a precursor of 217 amino acids. Furthermore, recombinant stGH was prepared and purified from Escherichia coli (E. coli) strain BL21. Using the 5×STAT5-Luciferase reporter system, we found that stGHR expressed in human hepatocellular carcinoma cells could be activated in a dose-dependent manner by recombinant stGH, thereby triggering the intracellular JAK2-STAT5 signaling pathway, suggesting stGH is a functional ligand for stGHR. Quantitative real-time PCR revealed that GH mRNA was predominantly and abundantly expressed in pituitary, while GHR exhibited ubiquitous expression in all tissues examined, including pituitary and liver. Collectively, our data indicate a conserved GH-GHR pair might function in reptiles including Chinese soft-shelled turtle, as that in mammals, birds and fishes.

To explore the temporal and spatial evolution of carbon sinks in state-owned forest regions (SOFRs) and the efficiency of increased carbon sinks, this study used panel data from 19 periods in 40 key SOFRs in Heilongjiang Province from 2001 to 2019. Additionally, combined with geographic information system (GIS) and remote sensing (RS) technology, the individual fixed-effect model was used to estimate the number of forest management investment (FMI) lagging periods, and the panel threshold model was used to investigate the differences in the FMI efficiency in various forest regions. From 2001 to 2019, the carbon sink of key SOFRs in Heilongjiang Province showed an upward trend over time, with a growth rate of 20.17%. Spatially, the phenomenon of “increasing as a whole and decreasing in a small area” was found, and the carbon sink of each forest region varied greatly. The standard deviation ellipse of the carbon sink presented a “southeast–northwest” pattern and had “from southeast to northwest” migration characteristics. The FMI amount from 2001 to 2019 showed an upward trend, with a total of CNY 46.745 billion, and varied greatly among forest regions. Additionally, the carbon sink amount in each SOFR affected the FMI efficiency. The threshold of the model was 5,327,211.8707 tons, and the elastic coefficients of the impact of FMI below and above the threshold on the carbon sink were 0.00953 and 0.02175, respectively. The latter’s FMI efficiency was 128.23% higher than that of the former. Finally, the increase in FMI to a carbon sink followed the law of diminishing marginal benefits. Therefore, the government should rationally plan the level of FMI in each SOFR to improve the FMI cost-effectiveness and help achieve the goal of “carbon neutrality”.

Malachite green oxalate (MG) is a kind of veterinary drug, which is freely soluble in water and hazardous to aquatic products, resulting in food toxicity and human health problems. The demand for effective and sensitive detection of MG residues is increasing in food safety. In this work, three DNA aptamers MG-36-12/16/17 targeting MG with good affinity (Kd values were 169.78, 71.94, and 102.46 μM, respectively) were obtained by Capture-SELEX. Furthermore, MG-36-12, MG-76-16-6A, and MG-36-17 were found to perform sensitively and specifically to detect MG as a sensing probe in a FRET fluorescent aptasensor, where the FAM-labeled aptamer and GO were employed as efficient energy donor–acceptor pair. The linear range of this aptasensor using aptamer MG-36-12 was from 1.71 to 514.29 ng/mL and the LOD was as low as 0.79 ng/mL. Additionally, the fluorescent assay using aptamer MG-36-17 to detect MG exhibited a linear relationship from 1.71 to 857.14 ng/mL and a LOD of 2.13 ng/mL. Meanwhile, the aptasensor showed high specificity to MG with no cross-reactivity to other veterinary drugs and had a mean recovery of 81.54% to 100.96% in actual water samples from the aquatic product market.

Software developers need information for deciding the optimal time for software release with improved software reliability. However, it is not easy for them to decide when and how to release newly developed software to the market. For a decision, the reliability and test costs of the software need to be balanced carefully for avoiding unnecessary confusion and users’ complaints. To address this need, related research has been carried out to propose an appropriate tool for such decisions. In many studies, software reliability growth models (SRGMs) were applied using the concept of confidence intervals to estimate the reliability of software. Confidence intervals were calculated on the basis of the assumption of a normal distribution showing the symmetrical occurrence of data with the mean as a center. However, the reliability data of software do not always have such symmetry for assuming the normal distribution. Therefore, it is necessary to propose a method for overcoming the mean value randomness that causes asymmetry in the related data. In previous studies, estimating variance and mean of errors of software was not considered, which led to the unreliable estimation of the confidence intervals of the mean value for decision making. Previous studies also lacked practicability in applications due to statistics from the asymmetrical data distribution. As a result, software developers could not effectively evaluate the possible risk related to the software release time. To improve the estimation, we employ the inflection S-shape model to propose the SRGM on the basis of confidence intervals assumed to come from the normal distribution. The proposed model allows determining the optimal time for software release with the consideration of its potential risk. For efficient determination, the architecture and user interface of the computation system are also proposed.

A cell surface displayed system in Pichia pastoris GS115 was developed by using GCW61, a glycosylphosphatidylinositol-modified cell wall protein from P. pastoris, as the anchor protein. Thermomyces lanuginosus lipase (TLL) was successfully displayed on the P. pastoris cell wall by fusing GCW61 gene with TLL2 gene (NCBI Accession: O59952) that was optimized with codon bias and synthesized. Cell surface displayed TLL2 was confirmed by the immunofluorescence microscopy. Flask fermentation was performed for 144 h with lipase activity up to 1964.76 U/g. Enzymatic properties of cell surface displayed TLL2 were also investigated. Displayed TLL2 occurred the maximum activity at pH 9 and 55°C and demonstrated characteristics of wide thermal adaptability and alkaline pH resistance. The optimum substrate was p-nitrophenyl hexanoate. Bivalent metal ions Ca2+, Mn2+, and Zn2+ had the activation effect on displayed TLL2, while Cu2+, Fe2+, Fe3+, K+, Li+, Na+, and Co2+ ions had the inhibitory effect on it. Since cell surface displayed TLL2 required less purification steps compared with free enzyme and showed high enzyme activities, it would be able to be further applied in various potential applications.

Objective: This study aimed to investigate the serum inflammatory cytokines levels in patients with COPD, pneumonia and lung cancer, and assess the correlation between the levels of inflammatory cytokines levels and development of these diseases. Methods: Two hundred thirty-two patients including 114 patients with pneumonia, 76 patients with chronic obstructive pulmonary disease (COPD) and 42 patients with lung cancer, and 62 age-matched healthy volunteers as controls were enrolled. The pro-inflammatory cytokine IL-6, IL-2, IFN-γ, TNF-α, anti-inflammatory cytokines IL-4 and IL-10 in serum were analyzed by flow cytometry microsphere array (CBA). Results: We found that the levels of TNF-α and IL-10 in patients with lung cancer, COPD and pneumonia were significantly higher than control group. The IL-6 in the lung cancer group were significantly increased compared with the controls and COPD group, pneumonia group. IFN-γ and IL-2 levels were lower in lung cancer compared with controls and COPD group, pneumonia group. TNF-α, IL-4 and IL-10 levels were increased in patients with COPD and pneumonia compared with controls. In addition, the concentrations of IFN-γ and IL-6 were increased in acute exacerbation COPD (AECOPD) group compared with stable COPD group. Conclusion: In conclusion, elevated TNF-α and IL-10 levels in serum may be related with lung diseases including lung cancer, COPD and pneumonia. Additionally, IFN-γ and IL-6 might be potential biomarkers for the further deterioration of lung disease patients. The increased concentrations of IFN-γ and IL-6 might be used to predict the exacerbation of COPD.

Acute leukemia is a common hematological malignancy. Despite recent promising progress, the prognosis of acute leukemia patients remains to be improved. New therapies are therefore still needed. Salmonella typhimurium has been shown to be highly effective as an anti-tumor agent in many solid cancer models, but it has not been applied in acute leukemia. Here, we report an attenuated Salmonella typhimurium strain, VNP20009, can induce apoptosis in multiple types of leukemia cells both in vivo and in vitro. Furthermore, VNP20009 significantly inhibited the proliferation of MLL-AF9-induced acute myeloid leukemia cells and prolonged the survival of the AML-carrying mice. VNP20009 restored the counts of white blood cell (WBC) and its five subsets in peripheral blood (PB) to near-physiological values, and elevated the levels of certain cytokines, such as tumor necrosis factor-α (TNF-α), leukemia inhibitory factor (LIF), interferon-γ (IFN-γ), chemokine C-X-C motif ligand-10 (CXCL-10) and C-C motif ligand-2 (CCL-2). Moreover, the ratio of immune cells, including natural killer cells (NKs), CD4+ Th1-type cells and CD8+ IFN-γ-producing effector T cells were highly upregulated in the AML mice treated with VNP20009. The results of the present study potentially provide an alternative therapeutic strategy for hematologic malignancies through boosting the innate and adaptive anti-tumor immunity.

Background: Cancer patients had been profoundly affected by the outbreak of COVID-19 especially after quarantine restrictions in China. We aimed to explore the treatment changes and delays of early breast cancer (EBC) during the first quarter of 2020. Methods: We did this retrospective, multicentre, cohort study at 97 cancer centres in China. EBC patients who received treatment regardless of preoperative therapy, surgery or postoperative therapy during first quarter of 2020 were included. Findings: 8397 patients were eligible with a median age of 50 (IQR 43–56). 0·2% (15/8397) of EBC patients were confirmed as COVID-19 infection. Only 5·2% of breast cancer diagnosis occurred after quarantine in Hubei compared with 15·3% in other provinces (OR= 0·30, 95%CI 0·24–0·38). postoperative endocrine therapy were least affected compared with different regions after quarantine (OR=0·37 [95%CI 0·19–0·73]). The proportion of surgery decreased from 16·4% in December last year to 2·6% in February in Hubei. Compared with intervals from diagnosis to treatment before quarantine restrictions, the average time increased with significance from 3·5 to 7·7 days in Hubei and 5·7 to 7·7 days in other provinces (p< 0·001). There were also 18·5 and 7·2 days delay in Hubei and other provinces respectively when calculating interval from surgery to postoperative therapy. Interpretation: EBC from high risk regions had a comparative rate of COVID-19 infection. After implementation of COVID-19 quarantine restrictions, fewer diagnosis and surgery with significant delays were seen when compared with treatment before. Funding: Beijing Medical Award Foundation (YJ0120)

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with a strong genetic basis. The role of de novo mutations in ASD has been well established, but the set of genes implicated to date is still far from complete. The current study employs a machine learning-based approach to predict ASD risk genes using features from spatiotemporal gene expression patterns in human brain, gene-level constraint metrics, and other gene variation features. The genes identified through our prediction model were enriched for independent sets of ASD risk genes, and tended to be down-expressed in ASD brains, especially in frontal and parietal cortex. The highest-ranked genes not only included those with strong prior evidence for involvement in ASD (for example, NBEA, HERC1, and TCF20), but also indicated potentially novel candidates, such as, MYCBP2 and CAND1, which are involved in protein ubiquitination. We also showed that our method outperformed state-of-the-art scoring systems for ranking curated ASD candidate genes. Gene ontology enrichment analysis of our predicted risk genes revealed biological processes clearly relevant to ASD, including neuronal signaling, neurogenesis, and chromatin remodeling, but also highlighted other potential mechanisms that might underlie ASD, such as regulation of RNA alternative splicing and ubiquitination pathway related to protein degradation. Our study demonstrates that human brain spatiotemporal gene expression patterns and gene-level constraint metrics can help predict ASD risk genes. Our gene ranking system provides a useful resource for prioritizing ASD candidate genes.

Background: Previous infectious or inflammatory events may be involved in the pathogenesis of neuromyelitis optica (NMO), potentially by triggering an autoimmune response. Cytomegalovirus (CMV)-related NMO (CMV-NMO) is rarely reported. Acute hemorrhagic rectal ulcer (AHRU) is a rare disease with a largely unknown pathogenesis. Herein, we reported a co-NMO and AHRU case associated with CMV infection. In addition, we review previously reported cases of CMV-NMO and CMV-AHRU.Case presentation: A 40-year-old female diagnosed with aquaporin4 (AQP4)-IgG+ NMO and a poor response to high-dose intravenous methylprednisolone and immunoglobulin, followed by three rounds of plasma exchange was transferred to Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. She developed repeated acute lower gastrointestinal hemorrhage from the third day of admission. Abdominal computed tomography angiography (CTA) and interventional angiography did not detect any bleeding vessel. Bedside colonoscopy revealed a large ulcer-like lesion at 10 cm above the anus. Rectal biopsy pathology confirmed a CMV infection on day 23 post-admission, and cerebrospinal fluid (CSF) pathogen gene sequencing detected CMV gene copies on day 25 post-admission. After 2 weeks of treatment with ganciclovir and sodium phosphinate, the patient's lower gastrointestinal bleeding stopped, and her limb muscle strength and visual acuity gradually improved. After 4 weeks of antiviral therapy, colonoscopy showed that the intestinal wall of the original lesion was smooth. Hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) of a biopsy specimen was negative for CMV, her right eye vision was normal, and limb muscle strength had recovered. Serum AQP4-IgG was negative, and lesions on brain magnetic resonance imaging (MRI) manifested shrinkage.Conclusions: The benefits of antiviral therapy remain unclear; however, clinicians should be aware of the possibility of CMV-related NMO, if NMO was refractory to high-dose intravenous methylprednisolone, immunoglobulin, and plasma exchange. Moreover, clinicians should consider the possibility of CMV-related AHRU when recurrent acute lower gastrointestinal bleeding occurs in a patient.

Pectate lyases play an essential role in textiles, animal feed, and oil extraction industries. Pichia pastoris can be an ideal platform for pectate lyases production, and BspPel (a thermo-alkaline pectate lyase from Bacillus sp. RN1) was overexpressed by combined strategies, reaching 1859 U/mL in a 50 L fermentator. It displayed the highest activity at 80°C, and maintained more than 60% of the activity between 30 and 70°C for 1 h. It showed an optimal pH of 10.0, and exhibited remarkable stability over a wider pH range (3.0-11.0), retaining more than 80.0% of enzyme activity for 4 h. The Km and kcat of BspPel on PGA (polygalacturonic acid) was 2.19 g L–1 and 116.1 s–1, respectively. The activity was significantly enhanced by Ca2+, Mn2+, and Cu2+, and a slight increase was observed with the addition of Ba2+ and Mg2+. Scanning electron microscope was used to show the degumming efficiency of BspPel on ramie fibers. The loss weight was 9.2% when treated with crude enzyme supernatant and 20.8% when treated with the enzyme-chemical method, which was higher than the 14.2% weight loss in the positive control treated with 0.5% (w/v) NaOH alone. In conclusion, BspPel could be a good candidate for the ramie degumming industry.

Abstract Background The non‐invasive prenatal screening (NIPS) has been introduced into clinical practice with a high sensitivity and specificity. Although the false‐negative results are inevitable and important, limited false‐negative NIPS results have been reported and studied previously. In this study, we aim to report and analyze false‐negative results detected in the NIPS cases with a low‐risk result. Methods NIPS was performed using whole‐genome massively parallel shotgun sequencing for screening common trisomies, rare autosomal aneuploidies, and subchromosome copy number variants. All the NIPS cases with a low‐risk result performed in our center in 2017 were followed‐up using medical records and telephone interview at 3 months after delivery. Fetal ultrasound results and available genetic diagnostic testing results were collected for pregnancies with adverse outcomes. The genetic diagnostic testing referred to chromosomal microarray analysis or fluorescent in situ hybridization on amniotic fluid cells, fetal skin tissue, neonatal peripheral blood, or available placental biopsies. Results By following‐up 10,975 low‐risk results, we found 166 NIPS cases with adverse pregnancy outcomes, in which eight cases had diagnostic testing. Among them, four false‐negative cases were confirmed, including one trisomy 18 caused by placental mosaicism, one mosaic tetrasomy 12p, and 2 microdeletion/microduplication cases. Conclusion Our results revealed that mosaicism contributes to a major cause of false negative in NIPS, and highlighted the importance of ultrasound in identifying these false‐negative results.

Element doping is an effective method to improve the performance of ZnO varistors. Previous studies mainly focused on the variation of microstructures and Schottky barriers. In this study, the effects of Co dopant on electrical properties are investigated from the aspect of multiscale defect structures, including intrinsic point defects, the heterogeneous interface of depletion/intergranular layers, and interface states at grain boundaries. Combining with analysis of phase composition and energy dispersive spectroscopy, it is found that Co tends to dissolve into ZnO grains when slightly doped. It substitutes Zn^2+ with the same valence and affects little on densities of donors. Segregation of Co at grain boundaries would result in the formation of spinel phase Co(Co(4/3)Sb(2/3))O(4) and transformation of the intergranular phase from α-Bi(2)O(3) to δ-Bi(2)O(3). Meanwhile, densities of point defects are indirectly affected by oxygen ambient during sintering, resulting in abnormal variation of grain resistivity. And interface states are enhanced, leading to improved barriers at grain boundaries. Therefore, reduced leakage current, enhanced grain resistivity, and improved non-linear coefficient in Co-doped ZnO varistor blocks are understood from the underlying multiple defect structures. This presents a potential approach to explore short-term performance and long-term stability of ZnO varistors from the aspect of defect responses.

Summary: Background: The Macrolides Oraux pour Réduire les Décès avec un Oeil sur la Résistance (MORDOR) trial found that biannual mass distribution of azithromycin to children younger than 5 years in Niger reduced the primary outcome of all-cause mortality by 18%. We aimed to determine the causes of mortality among deceased children using verbal autopsy. Methods: In this 2-year cluster-randomised controlled trial, 594 community clusters in Niger were randomly allocated (1:1 ratio) to receive biannual mass distributions of either oral azithromycin (approximately 20 mg per kg of bodyweight) or placebo targeted to children aged 1–59 months. Participants, study investigators, and field workers were masked to treatment allocation. Between Nov 23, 2014, and July 31, 2017, 3615 child deaths were recorded by use of biannual house-to-house censuses, and verbal autopsies were done between May 26, 2015, and May 17, 2018, to identify cause of death. Cause-specific mortality, as assessed by verbal autopsy, was a prespecified secondary outcome. This trial is completed and is registered with ClinicalTrials.gov, NCT02047981. Findings: Between Nov 23, 2014, and July 31, 2017, 303 communities (n=40 375 children at baseline) in Niger received mass azithromycin and 291 communities (n=35 747 children at baseline) received placebo. Treatment coverage was 90·3% (SD 10·6) in the azithromycin group and 90·4% (10·1) in the placebo group. No communities were lost to follow-up. In total, 1727 child deaths in the azithromycin group and 1888 child deaths in the placebo group were reported from the population censuses. Of these, the cause of death for 1566 (90·7%) children in the azithromycin group and 1735 (91·9%) children in the placebo group were ascertained by verbal autopsy interviews. In the azithromycin group, 437 (27·9%) deaths were due to malaria, 252 (16·1%) deaths were due to pneumonia, and 234 (14·9%) deaths were due to diarrhoea. In the placebo group, 493 (28·4%) deaths were due to malaria, 275 (15·9%) deaths were due to pneumonia, and 251 (14·5%) deaths were due to diarrhoea. Relative to communities that received placebo, child mortality in communities that received azithromycin was lower for malaria (incidence rate ratio 0·78, 95% CI 0·66–0·92; p=0·0029), dysentery (0·65, 0·44–0·94; p=0·025), meningitis (0·67, 0·46–0·97; p=0·036), and pneumonia (0·83, 0·68–1·00; p=0·051). The distribution of causes of death did not differ significantly between the two study groups (p=0·98). Interpretation: Mass azithromycin distribution resulted in approximately a third fewer deaths in children aged 1–59 months due to meningitis and dysentery, and a fifth fewer deaths due to malaria and pneumonia. The lack of difference in the distribution of causes of death between the azithromycin and placebo groups could be attributable to the broad spectrum of azithromycin activity and the study setting, in which most childhood deaths were due to infections. Funding: Bill & Melinda Gates Foundation.

ObjectiveDrawing a growth curve of multidrug-resistant Pseudomonas aeruginosa (MDR-PA) provides a foundation for susceptibility testing. By observing in vitro antibacterial activity and ultrastructure cthanges on MDR-PA of the effective components in the drug-containing serum of rats after the administration of Fuzheng Jiedu Huayu decoction (FJHD), we evaluated the inhibition and direct destruction effect of bacteria by TCM alone or combined with antibiotics.MethodsThe absorbance values of MDR-PA were determined at different detection time points, and a growth curve was drawn. After gavage with FJHD, drug-containing serum was collected from the rats. Using Imipenem/cilastatin sodium as the positive drug control, the in vitro antibacterial potency of FJHD and its drug-containing serum alone or in combination with antibiotics against MDR-PA was observed. The ultrastructural changes of MDR-PA treated by FJHD combined with antibiotics were observed by transmission electron microscopy.ResultsGrowth of the experimental strain manifested a lag phase in the first 1–4 h, an exponential growth phase at 5–20 h, and a plateau phase after 20 h. The best detection time during the susceptibility test was 16–20 h. The minimum inhibitory concentration (MIC) value of the FJHD extract group was 0.2 g/mL. The MIC value of the pure Imipenem/cilastatin sodium group was 16 μg/mL. The MIC values of Imipenem/cilastatin sodium + blank serum, 0.5-, 1-, and 2-fold drug-containing serum groups were all 16 μg/mL. The MIC values of Imipenem/cilastatin sodium + 4- and 8-fold drug-containing serum groups were both 8 μg/mL. By observation under a transmission electron microscope, Imipenem/cilastatin sodium + 0.5-, 1-, and 2-fold drug-containing serum groups showed bacterial structural damage. The degree of bacterial destruction was more obvious and the quantity of damaged bacteria was increased in the Imipenem/cilastatin sodium + 4- and 8-fold drug-containing serum groups.ConclusionDrawing the growth curve of the experimental strain had high application value for ensuring the accuracy of the drug sensitivity test results. TCM combined with antibiotics could enhance the antibacterial and direct destruction effect of bacteria in vitro, thereby inhibiting bacterial resistance to a certain extent.

Abstract Background Although tamoxifen is a highly effective drug for treating estrogen receptor–positive (ER+) breast cancer, nearly all patients with metastasis with initially responsive tumors eventually relapse, and die from acquired drug resistance. Unfortunately, few molecular mediators of tamoxifen resistance have been described. Here, we describe AFF3 (AF4/FMR2 family member 3), which encodes a nuclear protein with transactivation potential that confers tamoxifen resistance and enables estrogen-independent growth. Methods We investigated AFF3 expression in breast cancer cells and in clinical breast cancer specimens with western blot and Real-time PCR. We also examined the effects of AFF3 knockdown and overexpression on breast cancer cells using luciferase, tetrazolium, colony formation, and anchorage-independent growth assays in vitro and with nude mouse xenografting in vivo. Results AFF3 was overexpressed in tamoxifen-resistant tumors. AFF3 overexpression in breast cancer cells resulted in tamoxifen resistance, whereas RNA interference–mediated gene knockdown reversed this phenotype. Furthermore, AFF3 upregulation led to estrogen-independent growth in the xenograft assays. Mechanistic investigations revealed that AFF3 overexpression activated the ER signaling pathway and transcriptionally upregulated a subset of ER-regulated genes. Clinical analysis showed that increased AFF3 expression in ER+ breast tumors was associated with worse overall survival. Conclusions These studies establish AFF3 as a key mediator of estrogen-independent growth and tamoxifen resistance and as a potential novel diagnostic and therapeutic target.

Abstract Background The aim was to determine retinal nerve fiber layer function and its relations to retinal microvasculature and microcirculation in patients with myopia. Method Polarization-sensitive optical coherence tomography (PS-OCT) was used to measure phase retardation per unit depth (PR/UD, proportional to the birefringence) of the retinal nerve fiber layer (RNFL). Optical coherence tomography angiography (OCTA) was used to measure macular vessel density analyzed using fractal analysis. In addition, a retinal function imager (RFI) was used to measure macular blood flow velocities in arterioles and venules. Twenty-two patients with moderate myopia (MM, refraction > 3 and 0.05). The macular vessel density in both superficial and deep vascular plexuses was significantly lower in the HM group than in the other two groups (P 0.05). Conclusions The impairment of the retinal nerve fiber birefringence in the HM group may be one of the independent features in high myopic eyes, which appeared not to relate to macular microvascular density and blood flow velocity.