Publications by authors named "Zhixin Wang"

193 Publications

Apaf1 nanoLuc biosensors identified lentinan as a potent synergizer of cisplatin in targeting hepatocellular carcinoma cells.

Biochem Biophys Res Commun 2021 Aug 11;577:45-51. Epub 2021 Aug 11.

Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University, China. Electronic address:

Liver cancer is one of the most common malignancies that is difficult to treat due to late diagnosis and chemo-resistance. In the present study, we developed and validated a cell based split nanoLuc biosensor to monitor the Apaf1-Apaf1 interactions in response to apoptosis-inducing drugs such as cisplatin. We showed that the activity of split nanoLuc is reconstituted only in response to apoptotic inducer, cisplatin and in a dose-dependent manner. Apaf1 mutants which were unable to oligomerize failed to recover nanoLuc activity while constitutively active variant increased the nanoLuc activity. Generation of Apaf1 knockout HepG2 and treatment with cisplatin showed dramatic reduction in cell death suggesting that cisplatin mainly targets liver cancer cells through apoptosis. As the natural products are potent sources of compounds for adjuvant therapy, we screened a collection of natural products and identified lentinan as an inducer of apoptosome formation, a key step for induction of apoptosis. Lentinan is a polysaccharide with antitumor, pro-apoptotic properties that functions with poorly understood mechanisms. Lentinan was shown to have cytotoxic effects with the IC of 650 μM. Sub-lethal lentinan concentration doubled the nanoLuc activity when co-treated with cisplatin. We also showed that lentinan hugely reduced the dose of cisplatin to induce certain amount of death and that lentinan co-treatment with cisplatin enhanced the Apaf1 transcription in HepG2 cells while lentinan or cisplatin alone failed to alter the transcription. In addition, lentinan and cisplatin co-treatment induced mitochondrial depolarization. This suggested that lentinan combinatorial therapy with cisplatin engaged a different signalling pathway to kill the liver cancer cells and that adjuvant therapy with lentinan can reduce the dose of cisplatin and thus reduce the possibility of chemo-resistance.
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http://dx.doi.org/10.1016/j.bbrc.2021.08.030DOI Listing
August 2021

Metrological properties of neuropsychological tests for measuring cognitive change in individuals with prodromal Alzheimer's disease.

Aging Ment Health 2021 Aug 19:1-9. Epub 2021 Aug 19.

Department of Health Statistics, School of Public Health, Shanxi Medical University, Taiyuan, China.

Objectives: In Alzheimer's Disease (AD) research, choosing appropriate method for measuring change in cognitive function over time can be challenging. The aim for this study was to examine the sensitivity of four neuropsychological tests used to measure cognition during the transition from mild cognitive impairment (MCI) to AD, and the impacts of associated covariates.

Methods: We enrolled 223 patients with MCI who progressed to AD and had completed multiple follow-up assessments in the Alzheimer's Disease Neuroimaging Initiative (ADNI) database. We constructed nonlinear mixed model for multivariate longitudinal data assuming that multiple neuropsychological tests would exhibit nonlinear transformation of a common factor in the latent cognitive process underlying the progression from MCI to AD.

Results: The Clinical Dementia Rating-Sum of the Boxes (CDR-SB) and Alzheimer's Disease Assessment Scale (11 items; ADAS-11) were more sensitive to cognitive changes in individuals with higher cognitive function, the Functional Activities Questionnaire (FAQ) was more sensitive to cognitive changes in individuals with middle cognitive function, and the Mini-Mental State Examination (MMSE) was more sensitive to cognitive changes in individuals with lower cognitive function. Gender ( = 0.0139) and educational level ( = 0.0094) had varying effects on different tests, such that men performed better on the FAQ and CDR-SB, and individuals with higher educational level tended to perform better on the FAQ and MMSE.

Conclusions: When choosing appropriate neuropsychological tests in cognitive measurements, the cognitive functional level of the patient as well as the impacts of covariates should be considered.
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http://dx.doi.org/10.1080/13607863.2021.1966746DOI Listing
August 2021

Variations in Microbial Diversity and Metabolite Profiles of Female Landrace Finishing Pigs With Distinct Feed Efficiency.

Front Vet Sci 2021 9;8:702931. Epub 2021 Jul 9.

Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.

To enhance pig production, feed efficiency (FE) should be improved; however, the mechanisms by which gut microbes affect FE in pigs have not been fully elucidated. To investigate the differences between the composition and functionality of the gut microbiota associated with low and high FE, microbial compositions were characterized using 16S rRNA sequencing, functional annotations were performed by shotgun metagenomics, and metabolomic profiles were created by GC-TOF-MS from female Landrace finishing pigs with low and high feed conversion ratios (FCRs). was enriched in the gut microbiota of individuals with low FCRs (and thus high FE), while abundance was significantly higher in individuals with high FCRs (and thus low FE). This may be linked to carbohydrate consumption and incomplete digestion. The activity of pathways involved in the metabolism of cofactors and vitamins was greater in pigs with lower FE. We also identified differences in pyruvate-related metabolism, including phenylalanine and lysine metabolism. This suggests that pyruvate metabolism is closely related to microbial fermentation in the colon, which in turn affects glycolysis. This study deepens our understanding of how gut microbiota are related to pig growth traits, and how regulating microbial composition could aid in improving porcine FE. However, these results need to be validated using a larger pig cohort in the future.
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http://dx.doi.org/10.3389/fvets.2021.702931DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8299115PMC
July 2021

The Relationship Between Preoperative Systemic Immune Inflammation Index and Prognostic Nutritional Index and the Prognosis of Patients With Alveolar Hydatid Disease.

Front Immunol 2021 24;12:691364. Epub 2021 Jun 24.

Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University, Xining, China.

Background: To explore the relationship between the preoperative immune inflammation index (SII) and the prognostic nutritional index (PNI) and the overall survival rate (OS) of patients with alveolar hydatid disease.

Methods: The clinical data of patients with hepatic alveolar echinococcosis treated by surgery in the Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Qinghai University from January 2015 to January 2019 were analyzed retrospectively, and the SII, PNI, PLR and NLR were calculated. Spearman correlation analysis was utilized to analyze the correlation among SII, PNI, PLR and NLR. Receiver operating characteristic curve (ROC) was utilized to determine the best intercept values of SII, PNI, PLR and NLR, and Chi-square test was used to evaluate the relationship between SII, PNI and various clinicopathological features in patients with hepatic alveolar echinococcosis. The kaplan-Meier method was used to draw survival curves and analyze the relationship between them and the total survival time of patients. A cox regression model was used to analyze the relationship between SII, PNI and the prognosis of patients with hepatic alveolar echinococcosis. Finally, ROC curve was used to estimate the predictive efficacy of SII, PNI and COSII-PNI for the prognosis of patients with hepatic alveolar echinococcosis.

Results: A total of 242 patients were included, including 96 males and 146 females, aged 11.0-67.0 (36.6 ± 11.7) years. The values of SII, PNI, PLR and NLR are calculated, and the best truncation values of SII, PNI, PLR and NLR are given in ROC curve. The kaplan-Meier survival curve was used to analyze the relationship between SII, PNI, PLR, NLR and the overall survival time of patients with hepatic alveolar echinococcosis. The results showed that the median follow-up time was 45 months (95%CI: 39.484-50.516), and the average survival time was 49 months (95%CI: 47.300-51.931), which was low p<0.001); The 5-year OS rate of low PNI was significantly lower than that of high PNI group (37.7% 71.6%; p<0.001); The 5-year OS rate in low PLR group was significantly higher than that in high PLR group (70.4% 24.3%; p<0.001); The 5-year OS rate in low NLR group was significantly higher than that in high NLR group (67.2% 28.8%; p<0.001). Cox unifoliate analysis showed that SII, PNI, PLR and NLR were important prognostic factors related to OS. Cox multivariate analysis showed that SII(HR=4.678, 95% CI: 2.581-8.480, P<0.001) and PNI(HR=0.530, 95%CI: 0.305-0.920, P<0.05) were identified as independent risk indicators of OS, while NL was identified as independent risk indicators of OS ROC curve analysis showed that AUC of SII, PNI, PLR, NLR and COSII-PNI were 0.670(95%CI: 0.601-0.738), 0.638(95%CI: 0.561-0.716) and 0.618(95% CI: 0.541-0.694), respectively COSII-PNI is superior to SII and PNI in evaluating prognosis (P < 0.05).

Conclusions: SII and PNI can be regarded as independent risk factors reflecting the prognosis of patients with hepatic alveolar echinococcosis. The lower SII and the higher PNI before operation, the better the prognosis of patients, and the combined application of SII and PNI before operation can improve the accuracy of prediction.
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http://dx.doi.org/10.3389/fimmu.2021.691364DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264590PMC
June 2021

Assessing Genetic Diversity and Estimating the Inbreeding Effect on Economic Traits of Inner Mongolia White Cashmere Goats Through Pedigree Analysis.

Front Vet Sci 2021 22;8:665872. Epub 2021 Jun 22.

College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

The purpose of this study was to discover the population structure and genetic diversity of Inner Mongolia White Cashmere goats (IMCGs) and demonstrate the effect of inbreeding on the live body weight (LBW), cashmere yield (CY), fiber length (FL), and fiber diameter (FD) of IMCGs. All data were collected from pedigree information and production performance records of IMCGs from 1983 to 2019. The population structure and genetic diversity were analyzed by Endog 4.8 software. Inbreeding coefficients were obtained by the pedigree package in R. Then, a linear regression model was used to analyze how inbreeding influences economic traits in IMCGs. Four levels of inbreeding coefficients ( ) were classified in this study, including = 0, 0< ≤ 6.25, 6.25< ≤ 12.5 and ≥12.5. Variance analysis was performed to determine whether inbreeding levels had a significant effect on economic traits in IMCGs. The proportions of rams and dams in IMCGs for breeding were relatively small, with values of 0.8 and 20.5%, respectively. The proportion of inbred animals in the entire population was high, with values up to 68.6%; however, the average inbreeding coefficient and relatedness coefficient were 4.50 and 8.48%, respectively. To date, the population has experienced 12 generations. The average generation interval obtained in the present study was 4.11 ± 0.01 years. The ram-to-son pathway was lowest (3.97 years), and the ewe-to-daughter pathway was highest (4.24 years). It was discovered that the LBW, CY, and FL increased by 3.88 kg, 208.7 g, and 1.151 cm, respectively, with every 1% increase in the inbreeding coefficient, and the FD decreased by 0.819 μm with every 1% increase in the inbreeding coefficient. Additionally, multiple comparison analysis indicated that when the inbreeding coefficient was higher than 6.25%, the LBW showed an obvious decreasing trend. The threshold value of inbreeding depression in the CY is 12.5%. However, inbreeding depression has not been observed in the FL and FD. Pedigree completeness needs to be further strengthened. The degree of inbreeding in this flock should be properly controlled when designing breeding programs.
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http://dx.doi.org/10.3389/fvets.2021.665872DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8258104PMC
June 2021

Uncovering phylogenetic relationships and genetic diversity of water dropwort using phenotypic traits and SNP markers.

PLoS One 2021 6;16(7):e0249825. Epub 2021 Jul 6.

Institute of Vegetables, Wuhan Academy of Agricultural Sciences, Wuhan, Hubei, China.

The water dropworts Oenanthe linearis Wall. ex DC. and O. javanica (Blume) DC. are aquatic perennial herbs that have been used in China as vegetables and traditional medicines. However, their phylogenetic relationships and genetic diversity are poorly understood. Here, we presented the phenotypic traits and genome-wide DNA marker-based analysis of 158 water dropwort accessions representing both species. The analysis revealed that Oenanthe linearis was readily segregated into linear-leaf and deep-cleft leaf water dropworts according to their leaf shapes at flowering. Oenanthe javanica was classified by clustering analysis into two clusters based mainly on the morphological characteristics of their ultimate segments (leaflets). A set of 11 493 high-quality single-nucleotide polymorphisms was identified and used to construct a phylogenetic tree. There was strong discrimination between O. linearis and O. javanica, which was consistent with their phenotype diversification. The population structure and phylogenetic tree analyses suggested that the O. linearis accessions formed two major groups, corresponding to the linear-leaf and deep-cleft leaf types. The most obvious phenotypic differences between them were fully expressed at the reproductive growth stage. A single-nucleotide polymorphism-based analysis revealed that the O. javanica accessions could be categorized into groups I andII. However, this finding did not entirely align with the clusters revealed by morphological classification. Landraces were clustered into one group along with the remaining wild accessions. Hence, water dropwort domestication was short in duration. The level of genetic diversity for O. linearis (π = 0.1902) was slightly lower than that which was estimated for O. javanica (π = 0.2174). There was a low level of genetic differentiation between O. linearis and O. javanica (Fst = 0.0471). The mean genetic diversity among accessions ranged from 0.1818 for the linear-leaf types to 0.2318 for the groupII accessions. The phenotypic traits and the single-nucleotide polymorphism markers identified here lay empirical foundation for future genomic studies on water dropwort.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0249825PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8259969PMC
July 2021

High Co-Infection Rate of and Mycoplasma Girerdii in Gansu Province, China.

Healthcare (Basel) 2021 Jun 10;9(6). Epub 2021 Jun 10.

Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China.

() is a flagellated protist parasite that infects the human urogenital tract. The symbiotic relationship between and has been reported. Recent studies have identified a new strain, Mycoplasma girerdii ( M. girerdii), present in the vaginal secretions of women and have shown that this strain may be related to trichomoniasis. Here, we evaluated the presence of , and M. girerdii in 312 clinical samples from adult women diagnosed with vaginitis in Zhangye, Gansu province, China. Among these samples, 94, 153, and 48 were , and M. girerdii positive, respectively. Moreover, was highly frequent in 17-30-year-old women in this region. Forty samples (83.3%) positive for M. girerdii were also positive for . Six isolates were successfully cultured, including five isolates that showed symbiotic relationships with . This is the first report to evaluate the genetic characteristics of M. girerdii in China and may therefore provide insights into the effects of M. girerdii on the reproductive health of women.
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http://dx.doi.org/10.3390/healthcare9060706DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8227175PMC
June 2021

Mesophyll conductance variability of rice aquaporin knockout lines at different growth stages and growing environments.

Plant J 2021 Sep 2;107(5):1503-1512. Epub 2021 Aug 2.

National Key Laboratory of Crop Genetic Improvement, MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.

The plasma membrane subfamily of aquaporins [plasma membrane intrinsic proteins (PIPs)], which facilitates the CO diffusion across membranes, is proposed to play an important role in mesophyll conductance to CO (g ), a major limiting factor of photosynthesis. However, recent studies implied no causal relationship between g and PIPs because they failed to repeat the previous observed differences in g between PIP knockout lines and their wild-type. The contrasting results on the role of PIPs in g were interpreted as the different growth conditions among studies, which has never been tested. Here, we assessed whether the differences in g between wild-type and PIP knockout lines, Ospip1;1, Ospip1;2 and Ospip2;1, varied with growth condition (field versus pot condition) and growth stages in rice. Under field conditions, no differences were observed in plant performance, photosynthetic rate (A) and g between PIP knockout lines and the wild-type. However, in agreement with previous studies, two out of three knockout lines showed significant declines in tiller number, plant height, A and g under pot conditions. Moreover, we found that the differences in A and g between PIP knockout lines and the wild-type varied with the growth stage of the plants. Our results showed that the differences in g between PIP knockout lines and wild-type were depending on the growth environments and stage of the plants, and further efforts are required to reveal the underlying mechanisms.
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http://dx.doi.org/10.1111/tpj.15397DOI Listing
September 2021

Expression Profiling and Functional Analysis of Circular RNAs in Inner Mongolian Cashmere Goat Hair Follicles.

Front Genet 2021 11;12:678825. Epub 2021 Jun 11.

College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

Background: Inner Mongolian cashmere goats have hair of excellent quality and high economic value, and the skin hair follicle traits of cashmere goats have a direct and important effect on cashmere yield and quality. Circular RNA has been studied in a variety of tissues and cells.

Result: In this study, high-throughput sequencing was used to obtain the expression profiles of circular RNA (circRNA) in the hair follicles of Inner Mongolian cashmere goats at different embryonic stages (45, 55, 65, and 75 days). A total of 21,784 circRNAs were identified. At the same time, the differentially expressed circRNA in the six comparison groups formed in the four stages were: d75vsd45, 59 upregulated and 33 downregulated DE circRNAs; d75vsd55, 61 upregulated and 102 downregulated DE circRNAs; d75vsd65, 32 upregulated and 33 downregulated DE circRNAs; d65vsd55, 67 upregulated and 169 downregulated DE circRNAs; d65vsd45, 96 upregulated and 63 downregulated DE circRNAs; and d55vsd45, 76 upregulated and 42 downregulated DE circRNAs. Six DE circRNA were randomly selected to verify the reliability of the sequencing results by quantitative RT-PCR. Subsequently, the circRNA corresponding host genes were analyzed by the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The results showed that the biological processes related to hair follicle growth and development enriched by GO mainly included hair follicle morphogenesis and cell development, and the signaling pathways related to hair follicle development included the Notch signaling pathway and NF-κB signaling pathway. We combined the DE circRNA of d75vsd45 with miRNA and mRNA databases (unpublished) to construct the regulatory network of circRNA-miRNA-mRNA, and formed a total of 102 pairs of circRNA-miRNA and 126 pairs of miRNA-mRNA interactions. The binding relationship of circRNA3236-chi-miR-27b-3p and circRNA3236-chi-miR-16b-3p was further verified by dual-luciferase reporter assays, and the results showed that circRNA3236 and chi-miR-27b-3p, and circRNA3236 and chi-miR-16b-3p have a targeted binding relationship.

Conclusion: To summarize, we established the expression profiling of circRNA in the fetal skin hair follicles of cashmere goats, and found that the host gene of circRNA may be involved in the development of hair follicles of cashmere goats. The regulatory network of circRNA-miRNA-mRNA was constructed and preliminarily verified using DE circRNAs.
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http://dx.doi.org/10.3389/fgene.2021.678825DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8226234PMC
June 2021

Genome-guided investigation of anti-inflammatory sesterterpenoids with 5-15 trans-fused ring system from phytopathogenic fungi.

Appl Microbiol Biotechnol 2021 Jul 22;105(13):5407-5417. Epub 2021 Jun 22.

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

Fungal terpenoids catalyzed by bifunctional terpene synthases (BFTSs) possess interesting bioactive and chemical properties. In this study, an integrated approach of genome mining, heterologous expression, and in vitro enzymatic activity assay was used, and these identified a unique BFTS sub-clade critical to the formation of a 5-15 trans-fused bicyclic sesterterpene preterpestacin I (1). The 5-15 bicyclic BFTS gene clusters were highly conserved but showed relatively wide phylogenetic distribution across several species of the diverged fungal classes Dothideomycetes and Sordariomycetes. Further genomic organization analysis of these homologous biosynthetic gene clusters from this clade revealed a glycosyltransferase from the graminaceous pathogen Bipolaris sorokiniana isolate BS11134, which was absent in other 5-15 bicyclic BFTS gene clusters. Targeted isolation guided by BFTS gene deletion led to the identification of two new sesterterpenoids (4, and 6) from BS11134. Compounds 2 and 4 showed moderate effects on LPS-induced nitrous oxide production in the murine macrophage-like cell line RAW264.7 with in vitro inhibition rates of 36.6 ± 2.4% and 24.9 ± 2.1% at 10 μM, respectively. The plausible biosynthetic pathway of these identified compounds was proposed as well. This work revealed that phytopathogenic fungi can serve as important sources of active terpenoids via systematic analysis of the genomic organization of BFTS biosynthetic gene clusters, their phylogenetic distribution in fungi, and cyclization properties of their metabolic products. KEY POINTS: • Genome mining of the first BFTS BGC harboring a glycosyltransferase. • Gene-deletion guided isolation revealed three novel 5-15 bicyclic sesterterpenoids. • Biosynthetic pathway of isolated sesterterpenoids was proposed.
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http://dx.doi.org/10.1007/s00253-021-11192-3DOI Listing
July 2021

MicroRNA-mRNA Regulatory Networking Fine-Tunes Polyunsaturated Fatty Acid Synthesis and Metabolism in the Inner Mongolia Cashmere Goat.

Front Genet 2021 2;12:649015. Epub 2021 Jun 2.

College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

Fatty acid composition is an important aspect of meat quality in ruminants. Improving the beneficial fatty acid level in cashmere goat meat is important to its economic value. To investigate microRNAs (miRNAs) and mRNAs that regulate or coregulate polyunsaturated fatty acid (PUFA) synthesis and metabolism in the Inner Mongolia cashmere goat, we used longissimus dorsi muscle (WLM) and biceps femoris muscle (WBM) for transcript-level sequencing. RT-qPCR was used to evaluate the expression of mRNAs and miRNAs associated with PUFA synthesis and metabolism. The total PUFA content in the WBM was significantly higher than that in the WLM ( < 0.05). Our study is the first to systematically report miRNAs in cashmere goat meat. At the mRNA level, 20,375 genes were identified. , and were at the center of a gene regulatory network and contributed significantly to the accumulation and metabolic regulation of fatty acids. At the miRNA level, 426 known miRNAs and 30 novel miRNAs were identified. KEGG analysis revealed that the miRNA target genes were involved mainly in the PPAR signaling pathway. The mRNA-miRNA coregulation analysis showed that was negatively targeted by nine miRNAs: chi-miR-10a-5p, chi-miR-10b-5p, chi-miR-130b-5p, chi-miR-15a-5p_R-1, chi-miR-15b-5p, chi-miR-16a-5p, chi-miR-16b-5p, chi-miR-181c-5p_R+1, and chi-miR-26b-5p. Finally, we speculated that the simultaneous silencing of by one or more of these nine miRNAs through PPAR signaling led to low expression in the WLM and, ultimately to high PUFA content in the WBM. Our study helps elucidate the metabolic regulation of fatty acids in Inner Mongolia cashmere goats.
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http://dx.doi.org/10.3389/fgene.2021.649015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8206643PMC
June 2021

Fecal microbial composition and functional diversity of Wuzhishan pigs at different growth stages.

AMB Express 2021 Jun 12;11(1):88. Epub 2021 Jun 12.

State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.

The mammalian gut microbiome participates in almost all life processes in the host. In addition to diet, the breed is the main factor affecting changes in the swine gut microbiota. The composition of the gut microbiota changes significantly during different growth stages. Research on developmental changes in the gut microbiota of indigenous Chinese pig breeds is limited. In this study, the fecal microbiota of Wuzhishan pigs (a Chinese indigenous miniature pig) at different growth stages was investigated using high-throughput 16S rRNA sequencing. Firmicutes and Bacteroidetes were the two dominant phyla, accounting for more than 80% of all sequences. With increasing age, the fecal microbial diversity increased, and the proportion of Firmicutes increased, whereas the proportion of Bacteroidetes decreased. A total of 49 biomarkers with statistical differences were detected in the four growth stages. The different microbiota among groups enhanced the ability to degrade fiber, carbohydrates, and other substances during the growth stages. The endocrine system was different in multiple growth stage paired comparisons, which was attributed to the different body statuses in the growth stages. This study revealed developmental changes in the structure and function of gut microbes in local pigs.
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http://dx.doi.org/10.1186/s13568-021-01249-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8197691PMC
June 2021

The Correlation Between Platelet-to-Lymphocyte Ratio and Neutrophil-to-Lymphocyte Ratio with Hepatic Echinococcosis.

J Inflamm Res 2021 3;14:2403-2409. Epub 2021 Jun 3.

Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University, Xining, Qinghai, 810001, People's Republic of China.

Objective: To explore the correlation between neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) with hepatic hydatid diseases.

Methods: The clinical data of 420 patients with hydatid disease who were treated in our hospital from October 2015 to October 2018 were collected from the database of our hospital. According to the postoperative pathological diagnosis, 200 patients were assigned into the alveolar echinococcosis (AE) group, and 220 patients were assigned into the cystic echinococcosis (CE) group. A total of 160 healthy examinees were enrolled as the control group. The main observation indexes included preoperative PLR and NLR. Pearson's correlation was used to analyze the correlation between PLR and NLR with clinical indicators in HE patients, and the receiver operator characteristic (ROC) curve was used to evaluate the clinical values of PLR and NLR in diagnosing different types of hydatid diseases.

Results: The results revealed that the expressions of PLR and NLR were significantly higher in the AE group than in the CE and control groups (P<0.05). The results of multiple linear regression analysis revealed that PLR was positively correlated with aspartate aminotransferase, alanine aminotransferase, total bilirubin, direct bilirubin, alkaline phosphatase and γ-glutamyltranspeptidase (P<0.05). NLR was correlated with albumin content (P<0.05). ROC curve analysis results revealed that the area under the curve (AUC) of PLR and NLR for AE was 0.800 and 0.700 respectively, and the AUC for CE was 0.78 and 0.75 respectively.

Conclusion: The NLR and PLR of AE patients were higher than those of CE patients, and the high inflammation of CE patients may be correlated to the reproductive mode of . PLR and NLR have certain diagnostic values for disease classification, but PLR has higher specificity when compared with NLR.
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http://dx.doi.org/10.2147/JIR.S304307DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8184230PMC
June 2021

Genome-Based Discovery of Enantiomeric Pentacyclic Sesterterpenes Catalyzed by Fungal Bifunctional Terpene Synthases.

Org Lett 2021 Jun 4;23(12):4645-4650. Epub 2021 Jun 4.

State Key Laboratory of Bioreactor Engineering, East China University of Science of Technology, Shanghai 200237, China.

Genome-based discovery of two previously unreported fungal bifunctional terpene synthases (BFTSs) from phytopathogenic fungi are reported: FoFS catalyzing the formation of fusoxypenes A-C (-) and (-)-astellatene () and AtAS capable of synthesizing preaspterpenacid I (). Interestingly, FoFS and AtAS catalyzed the formation of enantiomeric sesterterpenes with a 5-6-7-3-5 ring system. C22-oxidative modification of preaspterpenacid I by AtP450 was characterized as well. Plausible cyclization pathways of the fusoxypenes were illustrated by DFT calculations.
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http://dx.doi.org/10.1021/acs.orglett.1c01361DOI Listing
June 2021

Pancreatic pseudocyst, pancreatitis, and incomplete pancreas divisum in a child treated with endotherapy: a case report.

J Int Med Res 2021 May;49(5):3000605211014395

Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Qinghai University, Xining, China.

Pancreatic divisum (PD) is caused by the lack of fusion of the pancreatic duct during the embryonic period. Considering the incidence rate of PD, clinicians lack an understanding of the disease, which is usually asymptomatic. Some patients with PD may experience recurrent pancreatitis and progress to chronic pancreatitis. Recently, a 13-year-old boy presented with pancreatic pseudocyst, recurrent pancreatitis, and incomplete PD, and we report this patient's clinical data regarding the diagnosis, medical imagining, and treatment. The patient had a history of recurrent pancreatitis and abdominal pain. Magnetic resonance cholangiopancreatography was chosen for diagnosis of PD, pancreatitis, and pancreatic pseudocyst, followed by endoscopic retrograde cholangiopancreatography, minor papillotomy, pancreatic pseudocyst drainage, and stent implantation. In the follow-up, the pseudocyst lesions were completely resolved, and no recurrent pancreatitis has been observed.
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http://dx.doi.org/10.1177/03000605211014395DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161893PMC
May 2021

Quantum Microwave Radiometry with a Superconducting Qubit.

Phys Rev Lett 2021 May;126(18):180501

Department of Applied Physics and Physics, Yale University, New Haven, Connecticut 06520, USA.

The interaction of photons and coherent quantum systems can be employed to detect electromagnetic radiation with remarkable sensitivity. We introduce a quantum radiometer based on the photon-induced dephasing process of a superconducting qubit for sensing microwave radiation at the subunit photon level. Using this radiometer, we demonstrate the radiative cooling of a 1 K microwave resonator and measure its mode temperature with an uncertainty ∼0.01  K. We thus develop a precise tool for studying the thermodynamics of quantum microwave circuits, which provides new solutions for calibrating hybrid quantum systems and detecting candidate particles for dark matter.
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http://dx.doi.org/10.1103/PhysRevLett.126.180501DOI Listing
May 2021

Genome Sequence Resource of Phytophthora colocasiae from China Using Nanopore Sequencing Technology.

Plant Dis 2021 May 13. Epub 2021 May 13.

Hainan University, 74629, Plant Pathology, 58 Renmin Avenue, Haikou, China, 570288.

Phytophthora colocasiae is a destructive oomycete pathogen of taro (Colocasia esculenta), which causes taro leaf blight. To date, only one highly fragmented Illumina short-read-based genome assembly is available for this species. To address this problem, we sequenced strain Lyd2019 from China using Oxford Nanopore Technologies (ONT) long-read sequencing and Illumina short-read sequencing. We generated a 92.51-Mb genome assembly consisting of 105 contigs with an N50 of 1.70 Mb and a maximum length of 4.17 Mb. In the genome assembly, we identified 52.78% repeats and 18,322 protein-coding genes, of which 12,782 genes were annotated. We also identified 191 candidate RXLR effectors and 1 candidate CRN effectors. The updated near-chromosome genome assembly and annotation resources will provide a better understanding of the infection mechanisms of P. colocasiae.
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http://dx.doi.org/10.1094/PDIS-11-20-2327-ADOI Listing
May 2021

Chi-miR-370-3p regulates hair follicle morphogenesis of Inner Mongolian cashmere goats.

G3 (Bethesda) 2021 05;11(5)

College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia, China.

MicroRNAs (miRNAs), a class of 22 nucleotide (nt) noncoding RNAs, negatively regulate mRNA posttranscriptional modification in various biological processes. Morphogenesis of skin hair follicles in cashmere goats is a dynamic process involving many key signaling molecules, but the associated cellular biological mechanisms induced by these key signaling molecules have not been reported. In this study, differential expression, bioinformatics, and Gene Ontology/Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on miRNA expression profiles of Inner Mongolian cashmere goats at 45, 55, and 65 days during the fetal period, and chi-miR-370-3p was identified and investigated further. Real-time fluorescence quantification (qRT-PCR), dual luciferase reporting, and Western blotting results showed that transforming growth factor beta receptor 2 (TGF-βR2) and fibroblast growth factor receptor 2 (FGFR2) were the target genes of chi-miR-370-3p. Chi-miR-370-3p also regulated the expression of TGF-βR2 and FGFR2 at mRNA and protein levels in epithelial cells and dermal fibroblasts. DNA staining, Cell Counting Kit-8, and fluorescein-labelled Annexin V results showed that chi-miR-370-3p inhibited the proliferation of epithelial cells and fibroblasts but had no effect on apoptosis. Cell scratch test results showed that chi-miR-370-3p promoted the migration of epithelial cells and fibroblasts. Chi-miR-370-3p inhibits the proliferation of epithelial cells and fibroblasts by targeting TGF-βR2 and FGFR2, thereby improving cell migration ability and ultimately regulating the fate of epithelial cells and dermal fibroblasts to develop the placode and dermal condensate, inducing hair follicle morphogenesis.
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http://dx.doi.org/10.1093/g3journal/jkab091DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8104936PMC
May 2021

Genome Sequence Resource of the Causal Agent of Stem and Root Rot of Cowpea.

Mol Plant Microbe Interact 2021 Aug 19:MPMI12200353A. Epub 2021 Aug 19.

Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, College of Plant Protection, Hainan University, Haikou 570228, China.

The causal agent of stem and root rot of cowpea, , is a widely distributed species of the genus. Here, we generate a high-quality complete genome assembly of PSY2020 (89.39 Mb, N 2.99 Mb) from China, using Oxford Nanopore Technologies (ONT) sequencing. The genome assembly completeness as evaluated by benchmarking universal single-copy orthologs was 94.51% at the eukaryote level. We identified 42.54% as repeat sequences and a total of 20,536 protein-encoding genes, of which 15,184 genes could be annotated. And we also identified 924 candidate RXLR effectors in the genome assembly. The described genome sequence will provide a valuable resource for better understanding of pathogenicity mechanisms of and help in uncovering phylogenetical classification of species.[Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 "No Rights Reserved" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. 2021.
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http://dx.doi.org/10.1094/MPMI-12-20-0353-ADOI Listing
August 2021

Review on the Adhesion of Geopolymer Coatings.

ACS Omega 2021 Mar 19;6(8):5108-5112. Epub 2021 Feb 19.

School of Civil Engineering, Binzhou University, Binzhou 256600, China.

Geopolymers are considered to be green materials with excellent fire resistance performance and potential substitutes for ordinary Portland cement (OPC). This review article focuses on the adhesion of geopolymer coatings subjected to elevated temperature. Their high adhesion strength is the basis for geopolymers being used as coating materials to work with the substrate. The adhesion strength is related to many factors, such as chemical composition of the raw materials, the formulation of the geopolymer, substrate type, surface roughness of the substrate, etc. The Si/Al ratio has different effects on compressive strength and bonding strength. The water content affects the polymerization process-the adhesion strength decreases with increasing water content. Careful tailoring of the mix ratio design is essential to make the geopolymer coating have excellent adhesive performance. These mix design factors include Si/Al ratio, Al/Na ratio, water content, precursor type, alkali cation type, curing conditions, etc.
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http://dx.doi.org/10.1021/acsomega.0c06343DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7931200PMC
March 2021

iTRAQ-based quantitative proteomic analysis of synergistic antibacterial mechanism of phenyllactic acid and lactic acid against Bacillus cereus.

Food Res Int 2021 01 1;139:109562. Epub 2020 Nov 1.

Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China. Electronic address:

Phenyllactic acid (PLA) as a phenolic acid by lactic acid (LA) bacteria shows enhanced antibacterial activity when coexisting with LA, while the antibacterial mechanism of PLA combined with LA was unknown. Hence, the antibacterial mechanism of PLA and LA was investigated against Bacillus cereus. Flow cytometry and TEM analysis demonstrated that single PLA and LA disrupted the membrane integrity and the morphology, while combined PLA and LA synergistically enhanced the damage. iTRAQ-based proteomic analysis suggested that PLA down-regulated kdpB and inhibited K transport, disturbed the function of ribosome and expression of competence genes; LA down-regulated periplasmic phosphorus-binding proteins and inhibited phosphorus transport, disturbed the function of ribosome, TCA cycle, as well as purine and pyrimidine metabolism; and combined PLA and LA inhibited K and phosphorus transport, and influenced the synthesis of purine and pyrimidine metabolism. The investigation could provide some insights into the application of PLA in food preservation.
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http://dx.doi.org/10.1016/j.foodres.2020.109562DOI Listing
January 2021

Genome Sequence Data of , an Oomycete Pathogen Causing Litchi Downy Blight.

Mol Plant Microbe Interact 2021 Jul 7:MPMI11200303A. Epub 2021 Jul 7.

Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, College of Plant Protection, Hainan University, Haikou 570228, China.

is an oomycete pathogen that exclusively infects litchi, with infection stages affecting a broad range of tissues. In this study, we obtained a near chromosome-level genome assembly of ZL2018 from China using Oxford Nanopore Technologies long-read sequencing and Illumina short-read sequencing. The genome assembly was 64.15 Mb in size and consisted of 81 contigs with an N of 1.43 Mb and a maximum length of 4.74 Mb. Excluding 34.67% of repeat sequences, 14,857 protein-coding genes were identified, among which 14,447 genes were annotated. We also predicted 306 candidate RxLR effectors in the assembly. The high-quality genome assembly and annotation resources reported in this study will provide new insight into the infection mechanisms of . [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 "No Rights Reserved" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. 2021.
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http://dx.doi.org/10.1094/MPMI-11-20-0303-ADOI Listing
July 2021

The intramolecular reaction of acetophenone -tosylhydrazone and vinyl: Brønsted acid-promoted cationic cyclization toward polysubstituted indenes.

Chem Commun (Camb) 2021 Feb;57(14):1810-1813

School of Petrochemical Engineering, and Jiangsu Key Laboratory of Advanced Catalytic Materials & Technology, Changzhou University, Changzhou 213164, P. R. China and Institute of New Materials & Industry Technology, College of Chemistry & Materials Engineering, Wenzhou University, Wenzhou 325035, P. R. China.

In the presence of TsNHNH2, a Brønsted acid-promoted intramolecular cyclization of o-(1-arylvinyl) acetophenone derivatives was developed, leading to polysubstituted indenes with complexity and diversity in moderate to excellent yields. In sharp contrast with either the radical or carbene involved cyclization of aldehydic N-tosylhydrazone with vinyl, a cationic cyclization pathway was involved, where N-tosylhydrazone served as an electrophile and alkylation reagent during this transformation.
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http://dx.doi.org/10.1039/d0cc07966aDOI Listing
February 2021

Localized increases in CEPT1 and ATGL elevate plasmalogen phosphatidylcholines in HDLs contributing to atheroprotective lipid profiles in hyperglycemic GCK-MODY.

Redox Biol 2021 04 6;40:101855. Epub 2021 Jan 6.

State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China; University of Chinese Academy of Sciences, Beijing, People's Republic of China. Electronic address:

Glucokinase-maturity onset diabetes of the young (GCK-MODY) represents a rare genetic disorder due to mutation in the glucokinase (GCK) gene. The low incidence of vascular complications in GCK-MODY makes it a natural paradigm for interrogating molecular mechanisms promoting vascular health under prolonged hyperglycemia. Clinical rate of misdiagnosis has remained high, and a reliable serum lipid biomarker that precedes genetic screening can facilitate correct diagnosis and treatment. Herein, we comprehensively quantitated 565 serum lipids from 25 classes in 105 subjects (42 nondiabetic controls, 30 GC K-MODY patients, 33 drug-naïve, and newly-onset T2D patients). At false-discovery rate (FDR) < 0.05, several phosphatidylcholines (PCs) and plasmalogen PCs were specifically increased in GCK-MODY, while triacylglycerols (TAGs) and diacylglycerols (DAGs) were reduced. Correlation matrices between lipids uncovered coregulation between plasmalogen PCs (PCps) and glycerolipid precursors was distinctly enhanced in GCK-MODY compared to T2D. Strengthened positive correlations between serum PCps and circulating HDLs was specifically observed in hyperglycemic subjects (i.e. T2D and GCK-MODY) compared to normglycemic controls, suggesting that HDL-PCps may elicit distinct physiological effects under hyperglycemia. Amongst GCK-MODY patients, individuals harboring variants of GCK mutations with elevated PCps also exhibited higher HDLs. Isolated HDLs displayed localized increases (p < 0.05) in very-long-chain PUFA-PCs and PCps in GCK-MODY. Protein analyses revealed elevated levels of HDL-resident ATGL (P = 0.003) and CEPT1 (P < 0.0001), which mediate critical steps of PCps production along the TAG-DAG-PC axis, in GCK-MODY relative to T2D. A panel of four lipids differentiated GCK-MODY from T2D with AUC of 0.950 (95% CI 0.903-9.997). This study provides the first evidence that enhanced recruitment of CEPT1 and ATGL onto HDLs essentially underlie the atheroprotective profiles associated with GCK-MODY. Resultant increases in the production of HDL-PCps and PUFA-PCs provides an active, circulating form of protection towards the vasculature of GCK-MODY, thereby lowering the incidence of vascular complications despite chronic exposure to hyperglycemia since birth.
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http://dx.doi.org/10.1016/j.redox.2021.101855DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7810764PMC
April 2021

Identification of abnormal protein expressions associated with mouse spermatogenesis induced by cyclophosphamide.

J Cell Mol Med 2021 02 12;25(3):1624-1632. Epub 2021 Jan 12.

Central Laboratory, Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China.

Cyclophosphamide (CP) is a clinical anticancer drug that can cause male reproductive abnormalities, but the underlying mechanisms for this remain unknown. The present study aimed to explore the potential toxicity induced by CP in spermatogenesis events of germ cell proliferation, meiosis, and blood-testis barrier integrity at the molecular level. CP-treated mice showed significantly reduced serum testosterone levels, sperm motility and concentration. The results of immunohistochemistry and Western blot showed that CP reduced the proliferation of germ cells (PCNA, PLZF) and increased germ cell apoptosis (Bax and TUNEL-positive cells) in CP-treated mice testes. The expression of meiotic related proteins (SYCP3, REC8, MLH1) decreased significantly in the fourth week after administration, and the expression of blood-testis barrier related proteins (β-catenin, ZO-1) and sperm quality-associated proteins (PGK2, HSPA4) decreased significantly in the first week after administration. CP leads to the apoptosis of male germ cells, inhibits the proliferation of germ cells, and affects meiosis and the blood-testis barrier, resulting in the decline of sperm quality. This study provides information to further the study of molecular mechanism and protective strategy of CP influence.
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http://dx.doi.org/10.1111/jcmm.16263DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7875923PMC
February 2021

DEPDC1 upregulation promotes cell proliferation and predicts poor prognosis in patients with gastric cancer.

Cancer Biomark 2021 ;30(3):299-307

Reproductive Medicine Centre, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong, China.

Background: Previous studies revealed that DEP domain containing 1 (DEPDC1) is involved in the carcinogenesis and progression of several types of human cancer. However the role of DEPDC1 in gastric cancer has not been studied.

Objective: The objective of this study was to study the expression and pathophysiological function of DEPDC1 in gastric cancer.

Methods: DEPDC1 expression in gastric adenocarcinoma cells was examined with Western blot and qRT-PCR. Clinical pathological features of patients were determined by immunohistochemistry. The effect of DEPDC1 expression on cell proliferation was studied by in vitro cell proliferation assay; and cell cycle influence was assessed by flow cytometry. Survival curves were plotted using Kaplan-Meier.

Results: DEPDC1 was overexpressed in gastric adenocarcinoma tissues compared with the paired adjacent normal gastric tissues, in accordance with mRNA level downloaded from GEPIA database. DEPDC1 expression level was significantly associated with cancer metastasis and differentiation. DEPDC1 upregulation caused cell cycle accelerating from G1 to S phase, and it was correlated with poorer overall survival.

Conclusion: Therefore, DEPDC1 upregulation in gastric adenocarcinoma is associated with tumor development and poor clinical outcomes of the patients, implying DEPDC1 might be a potential therapeutic target against gastric cancer.
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http://dx.doi.org/10.3233/CBM-201760DOI Listing
January 2021

Rapid Structure-Based Annotation and Profiling of Dihydrochalcones in Star Fruit () Using UHPLC/Q-Orbitrap-MS and Molecular Networking.

J Agric Food Chem 2021 Jan 23;69(1):555-567. Epub 2020 Dec 23.

Citrus Research & Education Center, Institute of Food and Agricultural Sciences, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850, United States.

Dihydrochalcones are a subclass of flavonoids. There has been growing interest in dihydrochalcones for their health benefits and potential to modulate flavor, but their comprehensive profile in diverse plant species is lacking. Star fruit is a tropical fruit rich in dihydrochalcones. In this study, a systematic annotation using UHPLC/Q-Orbitrap-MS and molecular networking was established to rapidly identify dihydrochalcones in 12 star fruit cultivars. A total of 53 dihydrochalcones were characterized within a short retention time including one novel compound (phloretin-3'-C-(2-O---coumaroyl)-β-d-fucopyranoside) and 23 compounds identified from the genus for the first time. 3-Hydroxyphloretin was the most abundant dihydrochalcone in star fruit. All the identified dihydrochalcones had a higher abundance in leaves compared to fruits. This is the first report that systematically investigates dihydrochalcones in star fruit of multiple cultivars, and the results could provide a useful reference for the future development and utilization of plant genetic resources.
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http://dx.doi.org/10.1021/acs.jafc.0c07362DOI Listing
January 2021

Genetic diversity and population structure of eddoe taro in China using genome-wide SNP markers.

PeerJ 2020 8;8:e10485. Epub 2020 Dec 8.

Wuhan Academy of Agricultural Sciences, Wuhan, Hubei, China.

Taro () is an important root and tuber crop cultivated worldwide. There are two main types of taro that vary in morphology of corm and cormel, 'dasheen' and 'eddoe'. The eddoe type () is predominantly distributed throughout China. Characterizing the genetic diversity present in the germplasm bank of taro is fundamental to better manage, conserve and utilize the genetic resources of this species. In this study, the genetic diversity of 234 taro accessions from 16 provinces of China was assessed using 132,869 single nucleotide polymorphism (SNP) markers identified by specific length amplified fragment-sequencing (SLAF-seq). Population structure and principal component analysis permitted the accessions to be categorized into eight groups. The genetic diversity and population differentiation of the eight groups were evaluated using the characterized SNPs. Analysis of molecular variance showed that the variation among eight inferred groups was higher than that within groups, while a relatively small variance was found among the two morphological types and 16 collection regions. Further, a core germplasm set comprising 41 taro accessions that maintained the genetic diversity of the entire collection was developed based on the genotype. This research is expected to be valuable for genetic characterization, germplasm conservation, and breeding of taro.
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http://dx.doi.org/10.7717/peerj.10485DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731653PMC
December 2020

Draft Genome of the European Mouflon ().

Front Genet 2020 19;11:533611. Epub 2020 Nov 19.

College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

Mouflon () with its huge and beautiful horns is considered as one of the ancestors of domesticated sheep. The European mouflon () is in the Asiatic mouflon () clade. In order to provide novel genome information for mouflon, moreover promote genetic analysis of genus both domestic and wild, we propose to sequence the mouflon genome. We assembled the highly heterozygous mouflon genome based on Illumina HiSeq platform using the next-generation sequencing technology. Finally, the draft genome we accessed approximately 2.69 Gb (42.15% GC), while N50 sizes of contig and scaffold are 110.1 kb and 10.4 Mb, respectively. The contiguity of this assembly is obviously better than earlier versions. Further analyses predicted 20,814 protein-coding genes in the mouflon genome and 12,390 shared gene families among bovine species. It is estimated that the divergence time between and was 7.6 million years ago. The draft mouflon genome assembly will provide data support and theoretical basis for various investigations of the genus species in future.
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http://dx.doi.org/10.3389/fgene.2020.533611DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7710762PMC
November 2020

Identification and biochemical characterisation of Acanthamoeba castellanii cysteine protease 3.

Parasit Vectors 2020 Nov 23;13(1):592. Epub 2020 Nov 23.

Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China.

Background: Acanthamoeba spp. are free-living amoeba that are ubiquitously distributed in the environment. This study examines pathogenic Acanthamoeba cysteine proteases (AcCPs) belonging to the cathepsin L-family and explores the mechanism of AcCP3 interaction with host cells.

Methods: Six AcCP genes were amplified by polymerase chain reaction (PCR). Quantitative real-time PCR was used to analyse the relative mRNA expression of AcCPs during the encystation process and between pre- and post-reactivated trophozoites. To further verify the role of AcCP3 in these processes, AcCP3 recombinant proteins were expressed in Escherichia coli, and the hydrolytic activity of AcCP3 was determined. The influence of the AcCP3 on the hydrolytic activity of trophozoites and the toxicity of trophozoites to human corneal epithelial cells (HCECs) was examined by inhibiting AcCP3 expression using siRNA. Furthermore, the levels of p-Raf and p-Erk were examined in HCECs following coculture with AcCP3 gene knockdown trophozoites by Western blotting.

Results: During encystation, five out of six AcCPs exhibited decreased expression, and only AcCP6 was substantially up-regulated at the mRNA level, indicating that most AcCPs were not directly correlated to encystation. Furthermore, six AcCPs exhibited increased expression level following trophozoite reactivation with HEp-2 cells, particularly AcCP3, indicating that these AcCPs might be virulent factors. After refolding of recombinant AcCP3 protein, the 27 kDa mature protein from the 34 kDa pro-protein hydrolysed host haemoglobin, collagen and albumin and showed high activity in an acidic environment. After AcCP3 knockdown, the hydrolytic activity of trophozoite crude protein against gelatin was decreased, suggesting that these trophozoites had decreased toxicity. Compared with untreated trophozoites or negative control siRNA-treated trophozoites, AcCP3-knockdown trophozoites were less able to penetrate and damage monolayers of HCECs. Western blot analysis showed that the activation levels of the Ras/Raf/Erk/p53 signalling pathways in HCECs decreased after inhibiting the expression of trophozoite AcCP3.

Conclusions: AcCP6 was correlated to encystation. Furthermore, AcCP3 was a virulent factor in trophozoites and participated in the activation of the Ras/Raf/Erk/p53 signalling pathways of host cells.
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http://dx.doi.org/10.1186/s13071-020-04474-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7685649PMC
November 2020
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