Comparative Transcriptome Analysis of Hepatopancreas Reveals Sexual Dimorphic Response to Methyl Farnesoate Injection in Litopenaeus vannamei.

Sexually dimorphic traits such as growth and body size are often found in various crustaceans. Methyl farnesoate (MF), the main active form of sesquiterpenoid hormone in crustaceans, plays vital roles in the regulation of their molting and reproduction. However, understanding on the sex differences in their hormonal regulation is limited. Here, we carried out a comprehensive investigation on sexual dimorphic responses to MF in the hepatopancreas of the most dominant aquacultural crustacean-the white-leg shrimp (Litopenaeus vannamei). Through comparative transcriptomic analysis of the main MF target tissue (hepatopancreas) from both female and male L. vannamei, two sets of sex-specific and four sets of sex-dose-specific differentially expressed transcripts (DETs) were identified after different doses of MF injection. Functional analysis of DETs showed that the male-specific DETs were mainly related to sugar and lipid metabolism, of which multiple chitinases were significantly up-regulated. In contrast, the female-specific DETs were mainly related to miRNA processing and immune responses. Further co-expression network analysis revealed 8 sex-specific response modules and 55 key regulatory transcripts, of which several key transcripts of genes related to energy metabolism and immune responses were identified, such as arginine kinase, tropomyosin, elongation of very long chain fatty acids protein 6, thioredoxin reductase, cysteine dioxygenase, lysosomal acid lipase, estradiol 17-beta-dehydrogenase 8, and sodium/potassium-transporting ATPase subunit alpha. Altogether, our study demonstrates the sex differences in the hormonal regulatory networks of L. vannamei, providing new insights into the molecular basis of MF regulatory mechanisms and sex dimorphism in prawn aquaculture.

The safety of maxillary sinus floor elevation and the accuracy of implant placement using dynamic navigation.

OBJECTIVE: To date, it remains a challenge to conduct maxillary sinus floor elevation (MSFE) owing to heterogeneity of anatomical structures and limited operative visibility of the maxillary sinus. The aim of this study is to investigate the safety of MSFE and the accuracy of implant placement using dynamic navigation. METHODS: Forty-two implants were placed in thirty-five patients requiring implantation in posterior maxilla with dynamic navigation. They were assigned to either lateral window sinus floor elevation (LWSFE) group (n = 22) or transcrestal sinus floor elevation (TSFE) group (n = 20) according to the residual alveolar bone height (RBH). Platform deviation, apex deviation and angular deviation between actual and planned implant placement were measured in precision evaluation software. Three deviations of two groups were compared via SPSS 22.0 software. RESULTS: Neither accidental bleeding nor perforation of Schneiderian membrane occurred in any patients. The actual window position of LWSFE was consistent with the preoperative design. There were no significant differences in platform, apex and angular deviations between the two groups (P > 0.05). CONCLUSION: In this study the dynamic navigation harvested clinically acceptable safety of MSFE and accuracy for implant placement in posterior maxillary region. The dynamic navigation would provide the clinician with assistance in achieving precise preoperative planning and reducing complications in surgical procedures. The granular bone grafts used in the LWSFE did not significantly affection on the accuracy of the simultaneous implant placement under the guidance of dynamic navigation.

A Convolutional Neural Network for Automated Detection of Cervical Ossification of the Posterior Longitudinal Ligament using Magnetic Resonance Imaging.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: We aimed to develop and validate a convolutional neural network (CNN) model to distinguish between cervical ossification of posterior longitudinal ligament (OPLL) and multilevel degenerative spinal stenosis using Magnetic Resonance Imaging (MRI) and to compare the diagnostic ability with spine surgeons. SUMMARY OF BACKGROUND DATA: Some artificial intelligence models have been applied in spinal image analysis and many of promising results were obtained; however, there was still no study attempted to develop a deep learning model in detecting cervical OPLL using MRI images. MATERIALS AND METHODS: In this retrospective study, 272 cervical OPLL and 412 degenerative patients underwent surgical treatment were enrolled and divided into the training (513 cases) and test dataset (171 cases). CNN models applying ResNet architecture with 34, 50, and 101 layers of residual blocks were constructed and trained with the sagittal MRI images from the training dataset. To evaluate the performance of CNN, the receiver operating characteristic curves of 3 ResNet models were plotted and the area under the curve were calculated on the test dataset. The accuracy, sensitivity, and specificity of the diagnosis by the CNN were calculated and compared with 3 senior spine surgeons. RESULTS: The diagnostic accuracies of our ResNet34, ResNet50, and ResNet101 models were 92.98%, 95.32%, and 97.66%, respectively; the area under the curve of receiver operating characteristic curves of these models were 0.914, 0.942, and 0.971, respectively. The accuracies and specificities of ResNet50 and ResNet101 models were significantly higher than all spine surgeons; for the sensitivity, ResNet101 model achieved better values than that of the 2 surgeons. CONCLUSION: The performance of our ResNet model in differentiating cervical OPLL from degenerative spinal stenosis using MRI is promising, better results were achieved with more layers of residual blocks applied.

Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan.

Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.

Molecular characterization and expression analysis of a QM protein gene in Chinese mitten crab Eriocheir sinensis.

QM protein was previously discovered as a tumor suppressor, and numerous studies have shown that QM protein also played important roles in the immune responses. To investigate the potential roles of the QM protein gene in Eriocheir sinensis, the QM protein gene (designated as EsQM) has been cloned from E. sinensis using the rapid amplification of cDNA ends (RACE) technique. The cDNA of EsQM is 781 bp in length, consisting of a 654 bp open reading frame (ORF), encoding 219 amino acids, a 27 bp 5' untranslated region (UTR) and a 94 bp 3' UTR. The EsQM protein has a calculated molecular weight of 25.4 kDa and a theoretical isoelectric point of 10.10. The deduced protein sequence of EsQM contains a Ribosomal_L16 domain, an SH3-binding motif, an N-acylation site, two putative antibiotic binding sites, two putative protein kinase C phosphorylation sites, and two amidation sites. EsQM is extremely conserved and exhibits more than 85% similarities to previously identified arthropod QM protein genes. By real-time quantitative PCR (qPCR) analysis, we found that EsQM mRNA transcripts were detectable in all the examined tissues, with the highest expression in hemocytes. The mRNA expression of EsQM in hemocytes was significantly upregulated after the stimulation of Aeromonas hydrophila or polybrominated diphenyl ether-47 (BDE-47). Moreover, EsQM mRNA expression in hemocytes responded more quickly and lasted longer when stimulated by A.hydrophila than BDE-47. Thus, EsQM can respond to bacterial infection and environmental pollution, and might be involved in the defense mechanism to both biological and non-biological stimulation of arthropods.

Urinary tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and cognitive functioning in older adults: The National Health and Nutrition Examination Survey 2013-2014.

INTRODUCTION: Tobacco contains carcinogens called tobacco-specific nitrosamines. Among the tobacco-specific nitrosamines, is nicotine-derived nitrosamine ketone (NNK) which produces the metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). We aimed to examine the association between urinary tobacco-specific NNAL and cognitive functioning among older adults. METHODS: A total of 1673 older adults aged ≥60 years from the National Health and Nutrition Examination Survey 2013-2014 were included. Urinary tobacco-specific NNAL was analyzed in the laboratory. Cognitive functioning was measured using the Consortium to Establish a Registry for Alzheimer's Disease Word Learning subtest (CERAD-WL) immediate and delayed memory tests, the Animal Fluency test (AFT), and the Digit Symbol Substitution Test (DSST). Test-specific and global cognition z-scores were calculated based on means and standard deviations of the cognitive test scores. Multivariable linear regression models were constructed to examine the independent association between quartiles of urinary tobacco-specific NNAL and cognitive test-specific and global cognition z-scores controlling for age, sex, race/ethnicity, education level, depressive symptoms, body mass index, systolic blood pressure, urinary creatinine, hypertension, diabetes, alcohol use, and smoking status. RESULTS: About half of the participants (mean age 69.8 years) were female (52.1%), non-Hispanic White (48.3%), and completed some college and above (49.7%). Multivariable linear regression results showed that participants in the 4th quartile (highest quartile) of urinary NNAL, compared with those in the 1st quartile (lowest quartile), had lower DSST z-scores (ß= -0.19; 95% CI: -0.34 - -0.04). CONCLUSIONS: Tobacco-specific NNAL was negatively associated with processing speed, sustained attention, and working memory in older adults.

Association between seropositivity for toxocariasis and cognitive functioning in older adults: an analysis of cross-sectional data from the US National Health and Nutrition Examination Survey (NHANES), 2011-2014.

OBJECTIVES: This study sought to examine the relationship between seropositivity for toxocariasis and cognitive functioning in a nationally representative sample of US older adults. DESIGN: A cross-sectional study. SETTING: National Health and Nutrition Examination Survey (NHANES) data collection took place in the US at participants' homes and mobile examination centres with specialised equipment. PARTICIPANTS: The study population consisted of 3188 community-dwelling US older adults aged 60 and above from the NHANES 2011 to 2014. OUTCOME MEASURES: IgG antibody against Toxocara spp was tested by a Luminex assay using recombinant rTc-CTL-1 antigen. A value >23.1 mean fluorescence intensity (MFI) indicated positive for toxocariasis and a value ≤23.1 MFI as negative for toxocariasis. The Consortium to Establish a Registry for Alzheimer's Disease Word Learning subtest immediate and delayed memory, the Animal Fluency test and the Digit Symbol Substitution Test (DSST) were used to assess cognitive functioning. Cognitive test-specific and global cognitive z scores were computed using sample means and SD. RESULTS: The study population consisted of 3188 participants who represented a total of 111 896 309 civilian citizens in the USA. The mean age of the participants was 69.6 years (standard deviation 6.8). The prevalence of toxocariasis in this population was 7.3% (95% confidence interval [CI] 6.1% to 8.5%). The survey-weighted linear regression model showed that compared with participants who were toxocariasis seronegative, those who were seropositive had lower DSST z score (beta [ß] = -0.12, 95% CI -0.22 to -0.01) and global cognition z score (ß=-0.11, 95% CI -0.22 to -0.01), after controlling for age, sex, race/ethnicity, education, depressive symptoms, smoking status, body mass index, prevalent coronary heart disease, prevalent stroke, and systolic blood pressure, physical activity, and total cholesterol. CONCLUSIONS: In our study, seropositive toxocariasis was independently and significantly associated with worse working memory, sustained attention, processing speed and global cognition in older adults. If this association is causal, public health measures to prevent human toxocariasis might help protect older adults' cognitive function.

Higher blood cotinine level is associated with worse cognitive functioning in non-smoking older adults.

Introduction: Secondhand smoke (SHS) is common in older adults; however, its cognitive effect is unclear. We aimed to examine the association between serum cotinine level and cognitive functioning among non-smoking older adults. Materials and methods: A total of 2,703 older adults aged 60 and above from the National Health and Nutrition Examination (NHANES) Survey 2011-2014 were included. Serum cotinine level was analyzed in the laboratory. A level ≤10 ng/ml and a response of "no" to the question "Do you currently smoke?" were used to select non-smokers. Cognitive functioning was measured using the Consortium to Establish a Registry for Alzheimer's disease Word Learning subtest (CERAD-WL) immediate and delayed recall tests, the Animal Fluency test (AFT), and the Digit Symbol Substitution test (DSST). Multivariable linear regression models were constructed to examine the association between serum cotinine level quartile and test-specific and global cognition z scores adjusting for age, race/ethnicity, education, depressive symptoms, body mass index, alcohol use, smoking history, prevalent coronary heart disease (CHD), stroke, and systolic blood pressure. Results: About half of the participants (mean age 70.5 years) were female (53.6%), non-Hispanic White (48.3%), and completed some college and above (50.2%). Multivariate linear regressions with a reference group being those in the 1st quantile (lowest) showed that participants in the 4th quartile (highest) of serum cotinine level had lower immediate recall [ß = -0.16, 95% confidence interval (CI) = -0.29, -0.03], AFT (ß = -0.19, 95% CI = -0.33, -0.05), DSST (ß = -0.27, 95% CI = -0.39, -0.15), and global cognition (ß = -0.26, 95% CI = -0.39, -0.14) z scores. Participants in the 3rd quartile had lower immediate recall (ß = -0.16, 95% CI = -0.30, -0.02) and global cognition (ß = -0.16, 95% CI = -0.29, -0.02) z scores. Participants in the 2nd quartile had lower delayed recall z scores (ß = -0.16, 95% CI = -0.29, -0.02). Conclusion: Higher serum cotinine level was associated with worse cognitive functioning in non-smoking older adults. Prevention and reduction of SHS in older adults may help protect their cognitive functioning.

Pre-clinical efficacy of CD20-targeted chimeric antigen receptor T cells for non-Hodgkin's lymphoma.

BACKGROUND: A 4-1BB/CD3-ζ-costimulated CAR-T against CD20 (CAR-T20) was subjected to a systemic efficacy evaluation in a cell co-culture model, and NOD-SCID IL-2 receptor gamma null mice (short for NSG mice) were xenografted with human Burkitt's lymphoma Raji cells. METHODS: CAR-T20 cells were incubated with target cells (K562, K562 CD20 or Raji cells) at ratios of 10:1 and 5:1 for 24 h, and the killing rate was estimated by an LDH cytotoxicity assay. To evaluate the effect of CAR-T20 on the survival time of tumor-bearing animals, 30 NSG mice were employed, and Raji-Luc cells (5 × 105 cells per mouse) were administered prior to CAR-T20 administration. The survival time, optical intensity of Raji-Luc cells, clinical symptoms, and body mass of the animals were observed. Another 144 male NSG mice were employed to investigate the proliferation and antitumor effects of CAR-T20. Human cytokine and murine cytokines were detected at 1, 7, 14, 21, 28, 42, 56 and 90 days post-CAR-T administration, while biochemistry index analysis, T-cell and CAR-T-cell detection in peripheral blood, and histopathological examination were performed at 14, 28, 56 and 90 days post-administration. RESULTS: CAR-T20 cells had a specific killing effect on CD20-expressing cells in vitro. At a dose of 1 × 106 per mouse or above, CAR-T20 prolonged the median survival time from 14 days to more than 3 months, inhibited the proliferation of Raji cells in mice, and alleviated the clinical manifestations and weight loss caused by the Raji-Luc cell load. CAR-T20 at a dose of 2 × 106 per mouse or above inhibited the proliferation of Raji cells in mice for up to 111 days post-administration without recurrence. The numbers of T cells and CAR-T cells in the animals administered CAR-T20 increased significantly when Raji cells were markedly proliferated and subsequently decreased when Raji cells were predominantly inhibited. CAR-T20 increased human IFN-γ, murine TNF and murine IL-6 levels and decreased human IL-10 levels in tumor-bearing mice. The incidences of xenografted tumors in organs/tissues were also reduced effectively by CAR-T20. CONCLUSION: The effective dose of CAR-T20 in mice starts from 1 × 106 per mouse, equivalent to a clinical dose of 5 × 106/kg. Together, our data support the clinical translation of CAR-T20 for R/R B-cell NHL patients.

The association between Roussouly sagittal alignment type and risk for adjacent segment degeneration following short-segment lumbar interbody fusion: a retrospective cohort study.

BACKGROUND: To date, the influence of Roussouly type on development of adjacent segment degeneration (ASD) after lumber fusion is still not fully explored, and the current study is aimed to evaluate the effect of Roussouly type on development of radiological ASD after single-level lumber fusion, and to compare the Roussouly types and spinopelvic parameters among those with different degenerative patterns of ASDs on sagittal plane. METHODS: A retrospective review of 288 patients underwent L4/5 or L5/S1 single-level posterior interbody fusions between January 2016 and December 2018 with a minimum 2-year follow up was performed. Radiological ASDs were identified and divided into 3 groups according to different degenerative patterns of the cephalad adjacent level on sagittal plane, including the types of retrolisthesis (Group A), anterolisthesis (Group B), and axial disc space narrowing (Group C). Roussouly types and radiological measurements were compared among three groups and potential risk factors for ASD were evaluated. RESULTS: Radiological ASD was found in 59 (20.5%) cases, in which patients with Roussouly type-2 was the most common. While, on subgroup analysis among three ASD groups, Roussouly type-1 occupied the highest proportion in Group A, differ in Group B and Group C, both with Type-2 as the most common. Moreover, Group A had significantly lower pelvic tilt (PT), larger sacral slope (SS), and larger segmental angle (SA) than Group B and Group C, which showed a more anteverted pelvic in Group A. Multivariate regression analysis noted Roussouly type, preoperative PT, and ∆PI-LL as the independent risk factors for radiological ASD. CONCLUSION: Roussouly type was significantly associated with the development of radiological ASD; however, the Roussouly types and spinal pelvic parameters were varied among different sagittal degenerative patterns of ASD, which was important in restoring optimal lumbar sagittal alignments in initial surgery.

Preclinical safety assessment of toxicity and biodistribution of oncolytic virus HSV-1 expressing human PD-1 antibody in mice.

HSV-1/hPD-1 is composed of engineered herpes simplex virus type-1 and two inserted copies of the human PD-1 antibody sequence. It is a novel oncolytic virus product designed to cure malignancies. The objective of this study was to estimate its toxicity in mice. In the single-dose toxicity study, no mortality and abnormal symptoms were observed in animals injected with 4.0 × 107 pfu/mouse dose. In the repeat-dose toxicity study, HSV-1/hPD-1 in animals intramuscularly treated with 1.0 × 107, 2.0 × 107, or 4.0 × 107 pfu/mouse doses was well tolerated in terms of clinical observation, body weight, food consumption, hematology and biochemistry indexes, T lymphocyte counting, immune reaction, and organ weight, except for some histopathological changes, such as the irreversible degeneration of the sciatic nerve, which was considered related to the adopted administration route. Synchronously, a biodistribution study in mice was performed to examine whether HSV-1/hPD-1 could spread to the injection site, gonads, liver, lung, heart, mesenteric and inguinal lymph nodes, skin, dorsal root ganglia, and blood, and then be gradually eliminated. Thus, two safety dose levels-the maximum tolerance dose of 4.0 × 107 pfu/mouse and the no-observed-adverse-effect-level dose of 1.0 × 107 pfu/mouse-were determined to help design patients' dose regimens. Our research data have been successfully accepted for investigational new drug (IND) application in China.

Tuning the Exchange Bias Effect in 2D van der Waals Ferro-/Antiferromagnetic Fe3 GeTe2 /CrOCl Heterostructures.

The exchange bias effect is extremely expected in 2D van der Waals (vdW) ferromagnetic (FM)/antiferromagnetic (AFM) heterostructures due to the high-quality interface. CrOCl possesses strong magnetic anisotropy at 2D limit, and is an ideal antiferromagnet for constructing FM/AFM heterostructures to explore the exchange bias effect. Here, the exchange bias effect in Fe3 GeTe2 (FGT)/CrOCl heterostructures through both anomalous Hall effect (AHE) and reflective magnetic circular dichroism (RMCD) measurements is studied. In the AHE measurements, the exchange bias field (HEB ) at 3 K exhibits a distinct increase from ≈150 Oe to ≈450 Oe after air exposure, and such variation is attributed to the formation of an oxidized layer in FGT by analyzing the cross-sectional microstructure. The HEB is successfully tuned by changing the FGT/CrOCl thickness and the cooling field. Furthermore, a larger HEB of ≈750 Oe at 1.7 K in FGT/CrOCl heterostructure through RMCD measurements is observed, and it is proposed that the larger HEB in RMCD measurements is related to the distribution of uncompensated spins at the interface. This work reveals several intriguing phenomena of the exchange bias effect in 2D vdW magnetic systems, which paves the way for the study of related spintronic devices.

Brain-wide Cas9-mediated cleavage of a gene causing familial Alzheimer's disease alleviates amyloid-related pathologies in mice.

The pathology of familial Alzheimer's disease, which is caused by dominant mutations in the gene that encodes amyloid-beta precursor protein (APP) and in those that encode presenilin 1 and presenilin 2, is characterized by extracellular amyloid plaques and intracellular neurofibrillary tangles in multiple brain regions. Here we show that the brain-wide selective disruption of a mutated APP allele in transgenic mouse models carrying the human APP Swedish mutation alleviates amyloid-beta-associated pathologies for at least six months after a single intrahippocampal administration of an adeno-associated virus that encodes both Cas9 and a single-guide RNA that targets the mutation. We also show that the deposition of amyloid-beta, as well as microgliosis, neurite dystrophy and the impairment of cognitive performance, can all be ameliorated when the CRISPR-Cas9 construct is delivered intravenously via a modified adeno-associated virus that can cross the blood-brain barrier. Brain-wide disease-modifying genome editing could represent a viable strategy for the treatment of familial Alzheimer's disease and other monogenic diseases that affect multiple brain regions.

Specific and behaviorally consequential astrocyte Gq GPCR signaling attenuation in vivo with ißARK.

Astrocytes respond to neurotransmitters and neuromodulators using G-protein-coupled receptors (GPCRs) to mediate physiological responses. Despite their importance, there has been no method to genetically, specifically, and effectively attenuate astrocyte Gq GPCR pathways to explore consequences of this prevalent signaling mechanism in vivo. We report a 122-residue inhibitory peptide from ß-adrenergic receptor kinase 1 (ißARK; and inactive D110A control) to attenuate astrocyte Gq GPCR signaling. ißARK significantly attenuated Gq GPCR Ca2+ signaling in brain slices and, in vivo, altered behavioral responses, spared other GPCR responses, and did not alter astrocyte spontaneous Ca2+ signals, morphology, electrophysiological properties, or gene expression in the striatum. Furthermore, brain-wide attenuation of astrocyte Gq GPCR signaling with ißARK using PHP.eB adeno-associated viruses (AAVs), when combined with c-Fos mapping, suggested nuclei-specific contributions to behavioral adaptation and spatial memory. ißARK extends the toolkit needed to explore functions of astrocyte Gq GPCR signaling within neural circuits in vivo.

ADAM10 promotes the proliferation of ligamentum flavum cells by activating the PI3K/AKT pathway.

Ligamentum flavum hypertrophy (LFH) is an important cause of spinal canal stenosis and posterior longitudinal ligament ossification. Although a number of studies have focused on the mechanisms responsible for LFH, the cellular mechanisms remain poorly understood. The aim of the present study was to investigate the roles of differentially expressed genes (DEGs) in LFH, elucidate the mechanisms responsible for LFH and provide a potential therapeutic target for further studies. The GSE113212 dataset was downloaded from the Gene Expression Omnibus (GEO) database. The microarray data were analyzed and DEGs were obtained. Bioinformatics methods, such as Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment and protein­protein interaction (PPI) network analyses were used to obtain the key genes and signaling pathways. In addition, cells derived from hypertrophied ligamentum flavum were cultured, and the key genes and signaling pathways in ligamentum cells were identified through in vitro cell biology and molecular biology experiments. A total of 2,123 genes were screened as DEGs. Among these DEGs, 1,384 genes were upregulated and 739 genes were downregulated. The KEGG pathway analysis revealed that the DEGs were mainly enriched in the PI3K/AKT signaling pathway, and the PPI network analysis screened A disintegrin and metalloproteinase 10 (ADAM10) as a key gene. In vitro experimental verification revealed that ADAM10 promoted the proliferation of ligamentum flavum cells and led to the hypertrophy of the ligamentum by activating the PI3K/AKT pathway. On the whole, the in vitro experimental results suggested that ADAM10 promoted the proliferation of ligamentum flavum cells by activating the PI3K/AKT pathway, which may represent a pathogenic mechanism of LFH. The findings of the present study may provide a basis and direction for further studies on the cellular mechanisms of LFH and present a potential novel therapeutic target and clinical approach.

Horseshoe crab genomes reveal the evolution of genes and microRNAs after three rounds of whole genome duplication.

Whole genome duplication (WGD) has occurred in relatively few sexually reproducing invertebrates. Consequently, the WGD that occurred in the common ancestor of horseshoe crabs ~135 million years ago provides a rare opportunity to decipher the evolutionary consequences of a duplicated invertebrate genome. Here, we present a high-quality genome assembly for the mangrove horseshoe crab Carcinoscorpius rotundicauda (1.7 Gb, N50 = 90.2 Mb, with 89.8% sequences anchored to 16 pseudomolecules, 2n = 32), and a resequenced genome of the tri-spine horseshoe crab Tachypleus tridentatus (1.7 Gb, N50 = 109.7 Mb). Analyses of gene families, microRNAs, and synteny show that horseshoe crabs have undergone three rounds (3R) of WGD. Comparison of C. rotundicauda and T. tridentatus genomes from populations from several geographic locations further elucidates the diverse fates of both coding and noncoding genes. Together, the present study represents a cornerstone for improving our understanding of invertebrate WGD events on the evolutionary fates of genes and microRNAs, at both the individual and population level. We also provide improved genomic resources for horseshoe crabs, of applied value for breeding programs and conservation of this fascinating and unusual invertebrate lineage.

Preclinical efficacy and safety evaluation of interleukin-6-knockdown CAR-T cells targeting at CD19.

BACKGROUND: ssCART-19 cells with shRNA-IL-6 gene knockdown were subjected to a comprehensive safety evaluation, including efficacy, toxicity and biodistribution studies in NSG (PrkdcscidIL2rgtm1 /Bcgen) mice. METHODS: NSG mice were administered Raji-Luc and then singly dosed with ssCART-19 cells via intravenous infusion. ssCART-19 DNA fragments were quantified in different tissues by qPCR, and the optical intensity of Raji-Luc was determined for evaluate the efficacy of regular CAR-T and ssCART-19 cells. In toxicity study, clinical symptoms observation, body weight measurements, serum biochemical analysis, human cytokine detection, lymphocytes subsets quantification, necropsy and histopathological examination were performed. RESULTS: The ssCART-19 DNA was mainly concentrated in the liver within 3 hours, and was widely distributed in most of the organs/tissues for 4 weeks after administration. Chimeric antigen receptor gene modified T cells (CAR-Ts) were detected in the peripheral blood with a significant increase in number beginning at approximately 3 weeks. ssCART-19 administration resulted in increased of interferon-gamma (IFN-γ), tumor necrosis factor (TNF), interleukin-2 (IL-2), and IL-17A and decreased IL-10 and IL-6 levels. ssCART-19 inhibited the proliferation of Raji-Luc cells in tumor-bearing NSG mice, and reduced the incidence of lymphomas in the liver, kidneys and spleen. It alleviated clinical symptoms caused by tumor cell proliferation in treated animals. CONCLUSIONS: ssCART-19 prolongs the survival time of tumor-bearing mice without obvious risks of immunotoxicity and tumorigenicity. ssCART-19 DNA was found in the brains of treated animals, however no significant central nervous system toxicity was observed. These data were used to support an investigational new drug (IND) application of ssCART-19 for clinical trial in China.

Tuning the Sensitivity and Dynamic Range of Optical Oxygen Sensing Films by Blending Various Polymer Matrices.

In this work, eight different types of optical oxygen sensing films were prepared by impregnating indicator and matrix solution on the surface of a polypropylene microporous filter membrane. The polymer matrix of the sensing films was ethyl cellulose (EC), polymethyl methacrylate (PMMA), and their blends with different mixing ratios. Scanning electron microscopy (SEM), laser confocal microscopy, and fluorescence spectrometer were used to investigate the morphologies and optical properties of the sensing films. Phase delay measurements under different oxygen partial pressures (PO2) and temperatures were applied to investigate the analytical performances of the sensing film for gaseous O2 monitoring. Results show that the response time of all the sensing films was extremely fast. The sensitivities and dynamic ranges of the sensing films with the blended polymer matrix were separately decreased and increased as the EC/PMMA ratio decreased, and the S-V curve of the sensing films blended with equal content of EC and PMMA exhibited good linearity under different temperatures, showing a promising prospect in practical application.

Tissue cross-reactivity studies of CPGJ701 in humans, cynomolgus monkeys and Sprague-Dawley rats and correlation analysis with in vivo toxicity.

BACKGROUND: CPGJ701 is a recombinant humanized anti-human epidermal growth factor receptor-2 (HER2) monoclonal antibody-derivative of the cytotoxic agent maytansine (DM1) conjugate for the treatment of HER2-positive metastatic breast cancer. Tissue cross-reactivity (TCR) studies of CPGJ701 in a complete panel of normal human, cynomolgus monkey and Sprague-Dawley were performed to provide evidence for selecting animal species for use in preclinical toxicity studies and predicting primary target organs and clinical adverse drug reactions (ADRs). METHODS: TCR studies were carried out to evaluate the distribution of reactivity and the TCR of CPGJ701 in paraffin sections of 32 tissues and/or organs (such as the heart, lung, liver, and kidney) from at least three unrelated normal human, cynomolgus monkey and Sprague-Dawley rat donors. The TCR of CPGJ701was detected by one-step immunohistochemical method using 50 µg/mL biotin-labeled CPGJ701 as the primary antibody. Moreover, a negative biotin-labeled human IgG control group, a blank phosphate-buffered saline (PBS) control group, and a positive human breast cancer tissue control group were also used to exclude false positive and false negative results. The specific positive binding and distribution of reactivity of CPGJ701 were detected in the human breast cancer tissue and 32 tissues from normal humans, cynomolgus monkeys and Sprague-Dawley rats under a microscope. RESULTS: The TCR of CPGJ701 in humans and cynomolgus monkeys was highly consistent but showed some differences compared to the TCR of CPGJ701 in Sprague-Dawley rats. The binding of CPGJ701 to target tissues, such as the liver, adrenal gland, thyroid, fallopian tube, spinal cord and skin, was observed in humans and cynomolgus monkeys but not in Sprague-Dawley rats. Specific binding to the placenta was only found in Sprague-Dawley rats. The cell types to which CPGJ701 specifically bound, including epithelial cells, cardiomyocytes and nerve cells, were identical in humans, cynomolgus monkeys and rats. CONCLUSIONS: The TCR of CPGJ701 was in accord with the targeting characteristics of the humanized anti-HER2 monoclonal antibody. The consistency of CPGJ701 binding to human and cynomolgus monkey tissues indicated that the cynomolgus monkey is a relevant animal species for evaluating the preclinical safety of CPGJ701. The targeting (binding site) of CPGJ701 in Sprague-Dawley rats indicated that it is also a useful animal species for evaluating antibody-dependent toxicity and non-antibody-dependent toxicity. In conclusion, these TCR studies of CPGJ701 could provide information for selecting relevant animal species for nonclinical studies and predicting clinical ADRs.

Intra-Articular Injection of Tranexamic Acid on Perioperative Blood Loss During Unicompartmental Knee Arthroplasty.

BACKGROUND Tranexamic acid (TXA) is safe and effective in total knee arthroplasty (TKA) for the prevention of bleeding. However, the role of TXA during unicompartmental knee arthroplasty (UKA) remains unclear. This study aimed to compare operative blood loss in patients undergoing UKA treated with an intra-articular injection of TXA with controls undergoing UKA without TXA. MATERIAL AND METHODS The prospective study included 101 patients who underwent UKA between January 2014 to March 2018. All patients completed a preoperative routine examination and were randomized to the study group (n=54) and the control group (n-47). The study group was given an articular injection of TXA (1.5 g in 50 ml normal saline) after the fascia was closed; the control group was injected with the same volume of normal saline. Blood volumes were measured from the drainage tube of the two groups during 48 hours. Total blood loss, postoperative drainage, hidden blood loss, blood transfusion rates, postoperative hemoglobin values, indicators of coagulation function, and the rates of wound complications were recorded. RESULTS Total blood loss in the study group was 745.6±105.1 ml, total drainage volume was 353.9±79.5 ml, and the hidden blood loss was 391.7±80.5 ml, which were all significantly lower when compared with the control group (P<0.05). None of the patients in the two groups suffered complications of surgery. CONCLUSIONS Intra-articular injection of TXA significantly reduced the total blood loss in patients who underwent UKA and did not increase the rate of complications.