Study on the Compositional Analysis, Extraction Process, and Hemostatic and Anti-Inflammatory Activities of Cirsium japonicum Fisch. ex DC.-Cirsium setosum (Willd.) MB Extracts.

Cirsium japonicum Fisch. ex DC. (CF) and Cirsium setosum (Willd.) MB (CS) are commonly used clinically to stop bleeding and eliminate carbuncles. Still, CF is mainly used for treating inflammation, while CS favors hemostasis. Therefore, the present study used UHPLC-MS to analyze the main chemical constituents in CF-CS extract. We optimized the extraction process using single-factor experiments and response surface methodology. Afterward, the hemostatic and anti-inflammatory effects of CF-CS extract were investigated by determining the clotting time in vitro, the bleeding time of rabbit trauma, and the induction of rabbit inflammation using xylene and lipopolysaccharide. The study of hemostatic and anti-inflammatory effects showed that the CF-CS, CF, and CS extract groups could significantly shorten the coagulation time and bleeding time of rabbits compared with the blank group (p < 0.01); compared with the model group, it could dramatically inhibit xylene-induced ear swelling in rabbits and the content of TNF-α, IL-6, and IL-1ß in the serum of rabbits (p < 0.01). The results showed that combined CF and CS synergistically increased efficacy. CF-CS solved the problem of the single hemostatic and anti-inflammatory efficacy of a single drug, which provided a new idea for the research and development of natural hemostatic and anti-inflammatory medicines.

Chromosome-scale genome assemblies of Himalopsyche anomala and Eubasilissa splendida (Insecta: Trichoptera).

Trichoptera is one of the most evolutionarily successful aquatic insect lineages and is highly valued value in adaptive evolution research. This study presents the chromosome-level genome assemblies of Himalopsyche anomala and Eubasilissa splendida achieved using PacBio, Illumina, and Hi-C sequencing. For H. anomala and E. splendida, assembly sizes were 663.43 and 859.28 Mb, with scaffold N50 lengths of 28.44 and 31.17 Mb, respectively. In H. anomala and E. splendida, we anchored 24 and 29 pseudochromosomes, and identified 11,469 and 10,554 protein-coding genes, respectively. The high-quality genomes of H. anomala and E. splendida provide critical genomic resources for understanding the evolution and ecology of Trichoptera and performing comparative genomics analyses.

Seven new species and four new records of Psychomyiidae (Insecta, Trichoptera) from China.

Seven new species of the family Psychomyiidae Walker, 1852 are described and illustrated from China; they are Psychomyiashunisp. nov., Ps.mangshanensissp. nov., Ps.capricornissp. nov., Lypesagittalissp. nov., Paduniellafasciariasp. nov., Pa.sanyaensissp. nov., and Tinodesaviformissp. nov. The genus Lype is reported for the first time from mainland China. In addition, four psychomyiids are found to be new to the Chinese caddis fauna: Psychomyiaindra Malicky & Chantaramongkol, 1993; Paduniellaandamanensis Malicky, 1979; Pa.dendrobia Malicky & Chantaramongkol, 1993; and Tinodesgapbona Johanson & Oláh, 2008. Moreover, Psychomyiapolyacantha Li, Qiu & Morse, 2021 is reviewed and synonymized with Psychomyiaimamiah Malicky, 2020.

Construction and application of a QSRR approach for identifying flavonoids.

A quantitative structure retention relationship (QSRR) method was developed to identify flavonoid isomers auxiliary using an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method based on the linear relationships between the Ln(k') values of flavonoids and their hydrogen bonding energy (XAH) and dissolution energy (ES). Chromatographic separation was achieved with a Hypersil GOLD C18 (100 mm × 2.1 mm, 1.9 µm) column and Agilent SB-C18 (2.1 ×50 mm, 1.8 µm) column on a Dionex Ultimate 3000 RSLC chromatograph. Compounds were eluted isocratically using a mobile phase containing 0.1% formic acid/water solution and methanol at a ratio of 55:45 (v/v). Mass spectrometry was performed in the negative and positive ionization modes on a Thermo Fisher Q Exactive Orbitrap mass spectrometer equipped with an electrospray ionization interface. The established QSRR model was Ln(k') = 5.6163 + 0.0469ES - 0.0984XAH, with a determination coefficient (R2) of 0.9981, adjusted determination coefficient (adjR2) of 0.9976, and corrected root mean square error of 0.0682. The determination coefficient of the leave-one-out (LOO) cross-validation (Q2LOO) was 0.9976, and the cross-verification root mean square error was 0.0754. Simulated samples containing 7 flavonoids were used to validate the feasibility of the method. The classical method (UHPLC-MS/MS combined the CD software and the mzCloud, mzVault and Chemspider databases) was used to identify the seven flavonoids in the simulated samples. This classic identification strategy cannot provide accurate identification results, which provided multiple identification results for each compound in the simulated samples. On the basis of the results, the 7 flavonoids were accurately identified by the established QSRR model, and the reference standards were used to validate it. The relative error of retention time(RE(tR)) between the model calculation and experimental results was less than 10%. This method effectively complements and improves the classical methods, that UHPLC-MS/MS combined the CD software and the mass spectra databases were used to identify flavonoids identification.

Investigation of internal damage evolution in gneiss considering water softening.

In soft rock tunnels, there are often large deformations during construction, especially when the groundwater seepage and softens the surrounding rock. For achieving the purpose of studying the softening effect of water immersion on strength and stability of surrounding rock, 15 rock samples were selected for physical and mechanical tests under 5 conditions: natural state and free immersion for 1, 3, 6, and 9 months, and nuclear magnetic resonance technology(NMR) was also adopted to test the internal pore structure of specimens with different immersion durations, thus the micro structure features of the gneiss, such as the NMR relaxation time T2 spectrum distribution, porosity, and pore volume ratio of different pore sizes under water softening were then obtained. The NMR results shows that the longer the free immersion duration of the rock sample, the greater the porosity; at the same time, the number of micropores in the rock gradually decreases under the interaction of water and rock, and the mesopores increase slightly first and then decrease all the time. The number of macropores is gradually increasing. When the immersion duration is 6 months, the number of macropores begins to increase significantly, and the mechanical properties of the specimens begin to drop significantly. By 9 months, the proportion of macropores in the rock has reached 57.6%. The results showed that the number growth of macropores is the root cause of the macroscopic failure of rock sample. The study results have significance for on-site construction in water-rich areas.

Four New Species of the Genus Eoneureclipsis (Trichoptera: Psychomyiidae) from China Inferred from Morphology and DNA Barcodes.

Four new species of the genus Eoneureclipsis Kimmins, 1955 from China are described, illustrated, and diagnosed based on male genitalia: Eoneureclipsis jianfenglingensis sp. nov. from Hainan, E. foraminulatus sp. nov. from Guangxi, E. spinosus sp. nov. from Guangxi and Guangdong, and E. gei sp. nov. from Fujian. A dichotomous key to Chinese adult males of Eoneureclipsis is provided. A distribution map for all Eoneureclipsis species is also presented. The DNA barcodes (partial mtCOI sequences) of E. jianfenglingensis sp. nov., E. gei sp. nov., and E. hainanensis Mey, 2013 have been generated and compared with all existing sequences of Eoneureclipsis species.

Circ-ABCC4 contributes to prostate cancer progression and radioresistance by mediating miR-1253/SOX4 cascade.

Circular RNAs (circRNAs) exert pivotal functions in many malignancies. However, the roles of circ-ABCC4 in prostate cancer (PCa) radioresistance and progression remain largely unclear. Cell viability, proliferation, apoptosis, invasion, and radioresistance were evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, 5-ethynyl-2'-deoxyuridine, flow cytometry, transwell invasion, and colony formation assays. Tumor xenograft experiment was conducted to assess circ-ABCC4 role in xenograft growth in vivo. Dual-luciferase reporter assay was implemented to test the target relation of microRNA-1253 (miR-1253) and circ-ABCC4 or SRY-box transcription factor 4 (SOX4). Circ-ABCC4 enrichment was prominently raised in PCa tissue specimens and cells. Circ-ABCC4 depletion blocked PCa cell viability, proliferation, invasion, and radioresistance and triggered apoptosis. Circ-ABCC4 silencing aggravated irradiation-induced inhibitory effect on xenografts growth. miR-1253 was a downstream molecule of circ-ABCC4, and circ-ABCC4 depletion-mediated anti-cancer impacts in PCa cells were partly counteracted by decreasing miR-1253 abundance. miR-1253 targeted SOX4 mRNA, and miR-1253 blocked PCa cell malignant phenotypes partly by targeting SOX4. Circ-ABCC4 could enhance SOX4 abundance by absorbing miR-1253. Circ-ABCC4 exerted a pro-tumor activity by facilitating PCa cell viability, proliferation, invasion, and radioresistance and suppressing apoptosis.

New Mitogenome Features of Philopotamidae (Insecta: Trichoptera) with Two New Species of Gunungiella.

A total of 14 individuals of Philopotamidae, from China, were examined. Six species in four genera, including two new species of the genus Gunungiella, were recognized. Their COI barcode sequences were extracted, mitogenomes were sequenced, assembled and analyzed. All of these sequences were used to further reveal the phylogenetic relationships of the family Philopotamidae. In addition, two new species: Gunungiella wangi n. sp., Gunungiella flabellata n. sp. were described and illustrated.

Seabuckthorn polysaccharide combined with astragalus polysaccharide ameliorate alcoholic fatty liver by regulating intestinal flora.

Background: At present, the incidence of alcoholic fatty liver disease (AFLD) is increasing year by year, and numerous studies have confirmed that liver diseases are closely related to intestinal flora. Seabuckthorn and Astragalus membranaceus, as traditional Chinese medicine (TCM) with the homology of medicine and food, have good liver protection, and their polysaccharides can regulate the intestinal flora. Here, we studied the effects of HRP, APS and the combination of the two polysaccharides on the intestinal flora of AFLD mice, which provided scientific basis for the treatment of AFLD with the two polysaccharides. Materials and methods: Thirty Kunming (KM) mice were randomly divided into the control group (Con), the model group (Mod), the HRP treatment group (HRP), the APS treatment group (APS), and HRP+APS treatment group (HRP+APS), with six mice in each group. The AFLD model was constructed by continuous intragastric administration of 42% vol Niulanshan ethanol solution for 28 days, and the mice in each polysaccharide group were given corresponding drugs. The levels of AST, ALT, TC and TG in serum of mice were measured. 16S rRNA amplicon sequencing technique was used to determine the diversity and richness of intestinal flora, and the relative abundance of intestinal flora at phylum level and genus level of the mice in each group. Results: HRP, APS and HRP+APS could reduce the serum levels of AST, ALT, TC and TG in mice. In addition, HRP, APS and HRP + APS restored the diversity, relative abundance and community structure of intestinal mucosa bacteria in AFLD mice to a certain extent. Specifically, HRP, APS and HRP+APS remarkably decreased the ratio of Firmicutes to Bacteroidetes, and ultimately increased the abundance of beneficial bacteria and reduced the abundance of pathogenic bacteria. Conclusion: HRP, APS, and HRP+APS can improve the intestinal microecology of AFLD model mice, alleviate liver injury, and maintain normal intestinal function in different degrees.

The First Chromosome-level Genome Assembly of Cheumatopsyche charites Malicky and Chantaramongkol, 1997 (Trichoptera: Hydropsychidae) Reveals How It Responds to Pollution.

Trichoptera is a highly adapted group of freshwater insects. They are generally more sensitive to dissolved oxygen and water quality than most freshwater organisms, and this sensitivity allows them to be used as reliable biological indicators of water quality. At present, there exists no chromosome-level genome of a hydropsychid species. Cheumatopsyche charites Malicky & Chantaramongkol, 1997 can successfully survive and thrive in polluted streams where other caddisflies are infrequent, suggesting that they are tolerant to latent contamination. Here we report a high-quality chromosome-level genome assembly of C. charites generated combining PacBio long reads and Hi-C reads. We obtained a genome assembly of 223.23 Mb, containing 68 scaffolds with an N50 length of 13.97 Mb, and 155 contigs (99.67%) anchored into 16 pseudochromosomes. We identified 36.12 Mb (16.18%) of the genome as being composed of repetitive elements, identified 369 noncoding RNAs, and predicted 8,772 protein-coding genes (96.80% BUSCO completeness). Gene family evolution analyses identified 7,148 gene families, of which 41 experienced rapid evolution. The expanded gene families were shown to be involved in detoxification metabolism, digestive absorption, and resistance to viruses or bacteria. This high-quality genome provides a valuable genomic basis for the study of trichopteran evolution.

Comparative Mitogenomic Analyses of Hydropsychidae Revealing the Novel Rearrangement of Protein-Coding Gene and tRNA (Trichoptera: Annulipalpia).

Gene rearrangement of the mitochondrial genome of insects, especially the rearrangement of protein-coding genes, has long been a hot topic for entomologists. Although mitochondrial gene rearrangement is common within Annulipalpia, protein-coding gene rearrangement is relatively rare. As the largest family in Annulipalpia, the available mitogenomes from Hydropsychidae Curtis, 1835 are scarce, and thus restrict our interpretation of the mitogenome characteristic. In this study, we obtained 19 novel mitogenomes of Hydropsychidae, of which the mitogenomes of the genus Arctopsyche are published for the first time. Coupled with published hydropsychid mitogenome, we analyzed the nucleotide composition evolutionary rates and gene rearrangements of the mitogenomes among subfamilies. As a result, we found two novel gene rearrangement patterns within Hydropsychidae, including rearrangement of protein-coding genes. Meanwhile, our results consider that the protein-coding gene arrangement of Potamyia can be interpreted by the tandem duplication/random loss (TDRL) model. In addition, the phylogenetic relationships within Hydropsychidae constructed by two strategies (Bayesian inference and maximum likelihood) strongly support the monophyly of Arctopscychinae, Diplectroninae, Hydropsychinae, and Macronematinae. Our study provides new insights into the mechanisms and patterns of mitogenome rearrangements in Hydropsychidae.

A New Species of Limnephilus (Insecta: Trichoptera: Limnephilidae) from China, with Revision of the Genus Limnephilus on the Chinese Mainland.

Fifty individuals of Limnephilus from the Qinghai Province, China, were examined, and their COI barcode sequences were extracted and analyzed. Forty individuals of Limnephilus from the Insect Collection of Nanjing Agricultural University (ICNAU), China, were examined, and photos of the male genitalia of four Limnephilus species are here presented. The males, females, larvae, and pupae of a new species, Limnephilus deqianensis n. sp., associated via COI barcode sequences, are described and illustrated. Ecological photos of the male, pupal case, and the habitat of the new species L. deqianensis n. sp. are also provided. Five species groups containing all seventeen Chinese Limnephilus species are revised. Diagnoses, keys, and a distribution map of them are provided. All of the sequences have been uploaded to GenBank. All specimens are deposited in the ICNAU, Nanjing, Jiangsu Province, P. R. China.

Study on the technology of efficient extraction of eleutheroside E from Acanthopanax senticosus by green solvent DES.

OBJECTIVES: Acanthopanax senticosus (Rupr. & Maxim.) Harms is a medicinal and edible plant which is clinically used for the recovery and treatment of cardiovascular and central diseases. As a characteristic active pharmaceutical ingredient of Acanthopanax senticosus, eleutheroside E is the core of the therapeutic effect. Organic solvent extraction has low selectivity, low extraction rate, difficulty in separation and purification and safety risks. The purpose of this study was to extract the effective component of Acanthopanax senticosus with a new green solvent. METHODS: In this article, two kinds of deep eutectic solvents (DESs) (DES-1 and DES-2) were synthesised by heating and stirring methods. Eleutheroside E was extracted by ultrasonic extraction with two kinds of DES as extractants and quantitatively analysed by Orbitrap-tandem mass spectrometry (MS/MS). RESULTS: The main results showed that the initial polarity of the DES was similar to that of 60 to 80% ethanol and hydrogen bond donors were the main factors affecting the polarity of DES. In the test, the viscosity of DES was higher than that of ethanol, and even the addition of a small amount of water (10%) caused intermolecular hydrogen bond disruption and redistribution of the solvent, resulting in a significant decrease in solvent viscosity. The solvents in the test group were stable after standing at 5°C in the dark for 100 days. The extraction rate of eleutheroside E by DES solvent was 5-6 times higher than that by ethanol. DES-1 and DES-2 can efficiently extract eleutheroside E. CONCLUSION: This study developed a new method for the application of the green extraction of eleutheroside E with certain practical significance.

FLG Gene Mutation Up-regulates the Abnormal Tumor Immune Response and Promotes the Progression of Prostate Cancer.

BACKGROUND: Prostate Cancer (PCa) ranks sixth with regard to the cause of cancerinduced male diseases worldwide, and inflammation is closely associated with its morbidity, deterioration, and prognosis. Tumor Mutation Burden (TMB) is identified to be the most common biomarker for the prediction of immunotherapy. But it is still unclear about the relationship of gene mutations in PCa with TMB and immune response. OBJECTIVES: To study the relationship between gene mutation and anti-tumor immune response in the prostate cancer tumor microenvironment. METHODS: In the present work, the PCa somatic mutation data were collected from the International Cancer Genome Consortium (ICGC) and The Cancer Genome Atlas (TCGA) datasets. RESULTS: As a result, 8 genes with high mutation frequency, including TP53, PTEN, TTN, FLG, CTNNB1, SPOP, MUC16, and KMT2C, were discovered to be covered by 4 cohorts from the United States, Canada, the United Kingdom, and China. Overall, the FLG mutation was related to a greater TMB, which predicted the dismal prognostic outcome. Besides, the CIBERSORT algorithm and Gene Set Enrichment Analysis (GSEA) were adopted for analysis, which revealed that FLG mutation remarkably promoted immune response in the context of PCa and accelerated cancer development. To sum up, FLG shows a high mutation frequency in PCa, and is related to the increase in TMB, up-regulation of abnormal immune responses in tumors, and promotion of tumor progression. CONCLUSION: Therefore, it may be used as a biomarker to predict the abnormal immune responses and provide a therapeutic target for immunotherapy in the treatment of PCa.

Protective effect of flavonoids extract of Hippophae rhamnoides L. on alcoholic fatty liver disease through regulating intestinal flora and inhibiting TAK1/p38MAPK/p65NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The therapeutic properties of Hippophae rhamnoides L. were already known in ancient Greece as well as in Tibetan and Mongolian medicine. Modern studies have indicated that Hippophae rhamnoides L. fermentation liquid protected against alcoholic fatty liver disease (AFLD). However, the underlying mechanism of Hippophae rhamnoides L. flavonoids extract (HLF) treating AFLD remains elusive. AIM OF THE STUDY: This study aimed to investigate the hepatoprotective effect of HLF in mice with AFLD and the interaction between AFLD and gut microbiota. MATERIALS AND METHODS: Chemical constituents of HLF were analyzed by Liquid Chromatography-Ion Trap-ESI-Mass Spectrometry. The Hepatoprotective effect of HLF was evaluated in mice with AFLD induced by alcohol (six groups, n = 10) daily at doses of 0.1, 0.2, and 0.4 g/kg for 30 consecutive days. At the end of experiment, mice were sacrificed and the liver, serum and feces were harvested for analysis. The liver histological changes were observed by H&E staining and oil red O staining. Moreover, the alterations of fecal microflora were detected by 16S rRNA gene sequencing. The inflammatory related genes were determined by qRT-PCR and western blotting respectively. RESULTS: The results showed that the oral administration of HLF remarkably alleviated hepatic lipid accumulation by decreasing the levels of ALT, AST, TG and TC. The levels of TNF-α, TGF-ß, and IL-6 were also reduced after treatment with HLF. Meanwhile, the protein and mRNA expression of NF-kB p65, MAPK p38 and TAK-1 in the liver of mice with AFLD were all reduced by HLF compared with model group. Furthermore, the 16S rRNA gene sequencing analysis demonstrated that HLF treatment can help restore the imbalance of intestinal microbial ecosystem and reverse the changes in Fimicutes/Bacterodietes, Clostridiales, Lachnospiraceae, S24-7, and Prevotella in mice with AFLD. CONCLUSION: HLF can effectively ameliorate liver injury in mice with AFLD, and regulate the composition of gut microbiota. Its regulatory mechanism may be related to TAK1/p38MAPK/p65NF-κB pathway. This study may provide novel insights into the mechanism of HLF on AFLD and a basis for promising clinical usage.

The First Report of the Genera Abaria and Drepanocentron (Trichoptera: Xiphocentronidae) from China, with Descriptions of Two New Species.

The genera Abaria Mosely 1948 and Drepanocentron Schmid 1982 are recorded in China for the first time. In this study, two new species, Abaria herringbona sp. nov., from Guang-xi, and Drepanocentron fuxiensis sp. nov., from An-hui, are described and illustrated. Male genitalia of these two new species is distinguishable from those of other Abaria and Drepanocentron species. In addition, Melanotrichia attia Malicky & Chantaramongkol 1992 is a new record for the Chinese caddisfly fauna.

Five new species of Tinodes (Trichoptera: Psychomyiidae) from China.

Five new species of genus Tinodes Curtis 1834 from China are described, illustrated, and diagnosed based on distinctive characters of male genitalia, including Tinodes falcata n. sp., T. xuzhouorum n. sp., T. shunhuangshanensis n. sp., T. yangae n. sp., and T. lini n. sp. Tinodes falcata n. sp. differs from other members of Tinodes in having an unpaired falcate and strongly sclerotized phallic sheath process. Tinodes xuzhouorum n. sp. differs from T. charmi Malicky 2009 by the relative lengths of the superior appendages and the phallus, the position of the ejaculatory duct, and the angle formed by tergum IX and the superior appendages in lateral view. Tinodes shunhuangshanensis n. sp. can be distinguished from T. sartael Malicky 2017 by the phallic sheath process splitting into paired dorsal branches and an apically incised ventral branch. Tinodes yangae n. sp. differs from T. eris Malicky & Sangpradub 2001 by the dorsal phallic sheath process having extra paired vertical branches. Tinodes lini n. sp. differs from T. dactringa Johanson & Oláh 2008 by the phallic guide with a pair of closely attached dorsal branches.

New species of the genus Molanna Curtis, 1834 (Trichoptera, Molannidae) in China inferred from morphology and DNA barcodes.

The male adult of Molannatruncata Ge, Peng & Sun sp. nov. is described and illustrated based on material collected in Si-chuan, China. It could be diagnosed by the subtriangular superior appendages when viewed dorsally, and by the mesal appendages each having a slender thorn and inferior appendages with a tiny inner process. Based on morphology of genitalia, we provide a dichotomous key to adult males of Molanna from the Oriental region. The DNA barcodes (partial mtCOI sequences) of M.truncata sp. nov. are generated and compared with existing sequences of Molanna species from Oriental and Palearctic regions. The mean intraspecific divergence of Molanna was 1.58% with a maximum of 8.50% in M.moesta. The Automatic Barcode Gap Discovery (ABGD) analysis of Molanna inferred 9 OTUs and thresholds of interspecific divergence of 10%. Divergence of M.truncata sp. nov. haplotypes from all other Molanna haplotypes ranged from 10.1% to 18%. We discuss distribution and potential groups of species within the Oriental Molanna species based on morphology.

Description of adult female, larva, and pupa of Glossosoma (Lipoglossa) kamael Malicky 2012 (Trichoptera: Glossosomatidae) associated by COI sequences from China.

The adult female, larva, and pupa of Glossosoma (Lipoglossa) kamael Malicky 2012 from Qinghai Province, China, are described and illustrated. Molecular associations are based on COI sequences. The immature stages of the subgenus Lipoglossa are described for the first time. Biological and habitat information of G. kamael are presented.

Four new species of Psychomyia (Trichoptera: Psychomyiidae) from China, with re-descriptions of two species.

Four new species of the genus Psychomyia from China are described, illustrated and diagnosed: P. complexa n. sp. (Zhe-jiang), P. conoidea n. sp. (Zhe-jiang), P. obtorta n. sp. (Jiang-xi), and P. tripetala n. sp. (Jiang-xi). Two known species, P. martynovi Hwang 1957 and P. extensa Li, Sun Yang 1999 are re-described and re-illustrated, with new distributional data.