Single-Cell RNA Transcriptome Profiling of Liver Cells of Short-Term Alcoholic Liver Injury in Mice.

Alcoholic liver disease (ALD) is currently considered a global healthcare problem with limited pharmacological treatment options. There are abundant cell types in the liver, such as hepatocytes, endothelial cells, Kupffer cells and so on, but little is known about which kind of liver cells play the most important role in the process of ALD. To obtain a cellular resolution of alcoholic liver injury pathogenesis, 51,619 liver single-cell transcriptomes (scRNA-seq) with different alcohol consumption durations were investigated, 12 liver cell types were identified, and the cellular and molecular mechanisms of the alcoholic liver injury were revealed. We found that more aberrantly differential expressed genes (DEGs) were present in hepatocytes, endothelial cells, and Kupffer cells than in other cell types in alcoholic treatment mice. Alcohol promoted the pathological processes of liver injury; the specific mechanisms involved: lipid metabolism, oxidative stress, hypoxia, complementation and anticoagulation, and hepatocyte energy metabolism on hepatocytes; NO production, immune regulation, epithelial and cell migration on endothelial cells; antigen presentation and energy metabolism on Kupffer cells, based on the GO analysis. In addition, our results showed that some transcription factors (TFs) are activated in alcohol-treated mice. In conclusion, our study improves the understanding of liver cell heterogeneity in alcohol-fed mice at the single-cell level. It has potential value for understanding key molecular mechanisms and improving current prevention and treatment strategies for short-term alcoholic liver injury.

Widely Targeted Metabolomics Analysis of Dendrobium officinale at Different Altitudes.

In order to investigate the differences in the chemical composition of Dendrobium officinale at different altitudes, we collected Dendrobium officinale from different altitudes in Guizhou Province, China, and firstly determined its polysaccharide content by using a sulfuric acid-phenol color development method with reference to the Chinese Pharmacopoeia, and further determined its metabolites by using widely targeted metabolomics, and explored the differences in the chemical composition of Dendrobium officinale at different altitudes using multivariate statistical analysis. We found that the polysaccharide content was higher in plants growing at 1122 m, a total of 902 secondary metabolites were detected by wildly targeted metabolomics, and amino acids and their derivatives were more highly expressed at 1122 m, while other metabolites were more highly expressed at 835 m. Furthermore, we found that the phenolic acid compound nerugein was only present in plants at 835 m, and two lipid compounds, namely Lyso PE 20 : 4 and its isomer, were only present at 1122 m. Taken together, these results may provide a basis for the selection and clinical application of D. officinale cultivated at different altitudes.

Ethnobotanical study of the wild edible and healthy functional plant resources of the Gelao people in northern Guizhou, China.

INTRODUCTION: The Gelao people are a unique minority in Southwest China with a unique culture for the utilization of edible plants, including a large number of medicinal plants. They believe that at least 61 species are edible and have medicinal value. Ethnobotany research can reveal the local knowledge of the Gelao people regarding the traditional use of plants and the relationship between this minority and their living environment to help retain and pass on this traditional knowledge forever. METHODS: Edible wild plants and their applied ethnic knowledge were investigated in three counties in northern Guizhou. Gelao residents were the main informants, and literature search, village interviews, participatory observation and quantitative ethnobotany evaluation were used. RESULTS: A total of 151 species of wild plants in 67 families are collected and eaten by Gelao residents, among which 61 species were considered to have medicinal value, accounting for 40.4% of the total, and 43 were listed in the Chinese Pharmacopoeia. There were 57 plant species with fruits as their edible parts, which are consumed as snacks, followed by 54 species whose young seedlings and leaves are the edible parts, most of which are consumed cold or stir-fried. Other edible parts included roots or rhizomes (bulbs), flowers, whole plants, seeds, fruiting bodies and stems. There were two consumption modes: raw and cooked. Raw foods were mainly consumed as snacks, which mainly comprise fruits. Cooked foods were mainly vegetables consumed cold or stir-fried. Some plants were used as seasonings, infused wines, condiments and grains. The main medicinal functions were nourishing and reducing heatiness. Nourishing plants were mainly "shen" plants and Liliaceae, while plants able to reduce heatiness were mainly Asteraceae. Others functions included anti-hangover, anticancer and insecticidal. There were 38 species of important edible wild plants (CFSI > 500) in northern Guizhou, which had a high utilization rate. Houttuynia cordata Thunb. and Mentha suaveolens Ehrh. were the most representative edible wild plants in this area. The species, edible parts, edible categories, consumption modes and medicinal functions of edible wild plants in this area are diverse, and the traditional knowledge on their uses is rich. However, the number of wild plant species eaten by the informants and their related knowledge were positively correlated with age, which indicates that the rich traditional knowledge in this area is gradually disappearing with urbanization. CONCLUSIONS: The Gelao have a rich history of consuming wild plants. With the development of the social economy, the traditional knowledge passed from older generations is gradually being lost and its inheritance is facing great risks. This study collects, sorts and spreads this precious traditional knowledge, which is of great value to its protection and inheritance and fully demonstrates the value and importance of our work.

Ethnobotany of wild edible plants in multiethnic areas of the Gansu-Ningxia-Inner Mongolia junction zone.

INTRODUCTION: In recent years, research on wild edible plant resources has become increasingly popular. The Hassan Nature Reserve is a multiethnic area mainly composed of people belonging to the Han, Hui, and Mongolian groups. The utilization of edible wild plant resources in this area is extremely high. However, with the advancement of urbanization and the development of modern agricultural technology, these resources have been seriously damaged, and related traditional knowledge, such as that related to national medicine, has been lost. METHODS: Based on a literature study, interviews with village and community organizations, participatory observation, and quantitative evaluation of ethnobotanical resources, wild edible plants in the Gansu-Ningxia-Inner Mongolia junction zone, were investigated. RESULTS: The survey results showed that there were 53 species (varieties) of wild edible plants belonging to 24 families in this area. The Compositae and Liliaceae families were the most abundant, with 8 and 7 species, respectively. The young stems and leaves were the most edible parts of the plants, as observed for 17 species, followed by fruits (including young fruits), which were considered the edible part of 16 species. Other edible parts included the roots or rhizomes (bulbs), seeds, whole plants, skins, etc. The edible plants were consumed in two forms: raw and cooked; raw plants, mainly fruit, were typically consumed as snacks. The cooked foods mainly consisted of vegetables, with tender stems and leaves as the main food source. These components were also used as seasoning, in medicinal diets, and as an emergency food source in times of famine. Important (CFSI > 500) wild edible plants used in health care in the region include Mulgedium tataricum (L.) DC., Nostoc commune Vaucher ex Bornet & Flahault, Sonchus arvensis L., Taraxacum mongolicum Hand.-Mazz., Allium schoenoprasum L., Robinia pseudoacacia L., Hemerocallis citrina Baroni, Elaeagnus angustifolia L., Medicago sativa L., Ulmus pumila L., Stachys sieboldii Miq., and Toona sinensis (Juss.) M. Roem., and these plants had high utilization values and rates locally. CONCLUSION: In summary, the species of wild edible plants and their edible parts, categories, consumption forms and roles in health care in this area are diverse. The utilization of traditional knowledge is rich, and some wild plants have high development value.

Compatibility effects of ginseng and Ligustrum lucidum Ait herb pair on hematopoietic recovery in mice with cyclophosphamide-induced myelosuppression and its material basis.

BACKGROUND: Ginseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. METHODS: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1, Rg5, salidroside, and so on. RESULTS: G, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit-granulocyte macrophage, burst-forming unit-erythroid, colony-forming unit-erythroid, and colony-forming unit-megakaryocyte and amount of G2/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. CONCLUSION: These results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.

Studies of the effects and mechanisms of ginsenoside Re and Rk3 on myelosuppression induced by cyclophosphamide.

BACKGROUND: Ginsenoside Re (Re) is one of the major components of Panax ginseng Meyer. Ginsenoside Rk3 (Rk3) is a secondary metabolite of Re. The aim of this study was to investigate and compare the effects and underlying mechanisms of Re and Rk3 on cyclophosphamide-induced myelosuppression. METHODS: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide. Peripheral blood cells, bone marrow nucleated cells, and colony yield of hematopoietic progenitor cells in vitro were counted. The levels of erythropoietin, thrombopoietin, and granulocyte macrophage colony-stimulating factor in plasma were measured by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. The expression of apoptotic protein bcl-2, bax, and caspase-3 was detected by Western blotting. RESULTS: Both Re and Rk3 could improve peripheral blood cells, bone marrow nucleated cell counts, thymus index, and spleen index. Furthermore, they could enhance the yield of colonies cultured in vitro and make the levels of granulocyte macrophage colony-stimulating factor, erythropoietin, and thrombopoietin normal, reduce the ratio of G0/G1 phase cells, and increase the proliferation index. Finally, Re and Rk3 could upregulate the expression of bcl-2, whereas they could downregulate the expression of bax and caspase-3. CONCLUSION: Re and Rk3 could improve the hematopoietic function of myelosuppressed mice. The effect of Rk3 was superior to that of Re at any dose. Regulating the levels of cytokines, promoting cells enter the normal cell cycle, regulating the balance of bcl-2/bax, and inhibiting the expression of caspase-3 may be the effects of Re and Rk3 on myelosuppression.

The effects of Arctigenin-Valine ester on chemotherapy-induced myelosuppression in mice.

OBJECTIVE: To explore whether Arctigenin-Valine ester (ARG-V) can treat myelosuppression caused by chemotherapy. METHODS: The number of peripheral blood cells of the mice was measured by an automatic blood analyzer, and the hematopoietic progenitor colonies CFU-GM, CFU-E, BFU-E, and CFU-Meg were cultured in vitro. Hematopoietic progenitor colonies and BMNCs were counted under an inverted microscope. The expressions of cytokines GM-CSF, EPO and TPO were detected by ELISA. The cell cycle was measured by flow cytometry. The expressions of related proteins MEK and p-ERK were quantitated by western blots, and the thymus index and spleen index were quantitated. RESULTS: After taking ARG-V, the peripheral blood cells of the mice gradually returned to normal, the number of nucleated cells in the bone marrow increased, the thymus index increased, the spleen index decreased, the number of hematopoietic progenitor cells increased, and the hematopoietic cytokines decreased. And ARG-V promoted the transformation of myelosuppression cells from G0/G1 to S and from S to G2/M. ARG-V could up-regulate the expression of MEK and p-ERK, and low dose ARG-V is not as effective in all aspects as high dose ARG-V. CONCLUSION: ARG-V can effectively alleviate the myelosuppression that caused by intraperitoneal injection of CTX in 100mg/kg, and ARG-V can promote the proliferation and differentiation of hematopoietic progenitor cells and improve immunity, and the effect of high-dose Arctigenin-Valine ester is more significant to some extent.

Comparative analysis of active ingredients and effects of the combination of Panax ginseng and Ophiopogon japonicus at different proportions on chemotherapy-induced myelosuppression mouse.

In this study, we aimed to investigate the effects of the combination of Panax ginseng and Ophiopogon japonicus (PG-OJ) herbs at different ratios on myelosuppression induced by chemotherapy. The myelosuppression model was established using an intraperitoneal injection of 100 mg kg-1 cyclophosphamide (CTX) in mice. The mice were administered the PG-OJ extract or Shengmaiyin (SMY) at different proportions (1 : 0, 1 : 1, 1 : 2, 2 : 1, 2 : 3, 3 : 2, and 0 : 1). The changes in the chemical composition caused by decocting the herbs together were analyzed by HPLC. The parameters i.e. the number of bone marrow nucleated cells and peripheral blood cells and the thymus and spleen indices were determined after administration. The results indicated that the co-decoction of PG and OJ, especially at the ratio of 2 : 3, was more conducive to the conversion of conventional ginsenosides (Rg1, Re, Rb1, Rg2, Rc, Rb2, Rb3 and Rd) to rare ginsenosides (Rg5, Rk3, S-Rg3, R-Rg3, Rk1 and Rh1) and the dissolution of ophiopogonin D. In addition, PG-OJ has an excellent synergistic effect on myelosuppression induced by CTX in mice. PG-OJ could significantly increase the numbers of the bone marrow nucleated cells and peripheral blood cells; moreover, it increased the thymus index and decreased the spleen index. The herb pair with a ratio of 2 : 3 showed the best therapeutic effect. By combining the results of the chemical composition changes and pharmacological activities, it can be concluded that rare ginsenosides and ophiopogonin D may be the main material basis of PG-OJ for the treatment of bone marrow suppression after chemotherapy.

A circular RNA circ_0000115 in response to graphene oxide in nematodes.

Circular RNAs (circRNAs) play important roles in regulating various biological processes; however, their roles in regulating the toxicity of engineered nanomaterials (ENMs) are still unclear. Based on Illumina HiSeq2500 sequencing, we here identified 43 dysregulated circRNAs in graphene oxide (GO) (1 mg L-1) exposed nematodes. Five of these candidate circRNAs could be further dysregulated by GO exposure in the range of µg L-1. Using the RNA interference (RNAi) technique, we found that the alteration in expressions of circ_0000115, circ_0000247, and circ_0000665 mediated a protective response to GO exposure; however, the alteration in expressions of circ_0000201 and circ_0000308 mediated the toxicity induction of GO. In nematodes, the circ_0000115 acted in certain tissues (intestine and neurons) to regulate GO toxicity. Moreover, an intermediate filament protein IFC-2 required for intestinal development was identified as a target of circ_0000115 in regulating the GO toxicity. In the intestine, intestinal IFC-2 acted further upstream of FOXO transcriptional factor DAF-16 in the insulin signaling pathway to regulate the GO toxicity. Therefore, intestinal circ_0000115 in the signaling cascade of circ_0000115-IFC-2-DAF-16 regulates the GO toxicity by modulating the function of IFC-2.

Institute collection and analysis of Nanobodies (iCAN): a comprehensive database and analysis platform for nanobodies.

BACKGROUND: Nanobodies are single-domain antibodies that contain the unique structural and functional properties of naturally-occurring heavy chain in camelidae. As a novel class of antibody, they show many advantages compared with traditional antibodies such as smaller size, higher stability, improved specificity, more easily expressed in microorganisms. These unusual hallmarks make them as promising tools in basic research and clinical practice. Although thousands of nanobodies are known to be published, no single database provides searchable, unified annotation and integrative analysis tools for these various nanobodies. RESULTS: Here, we present the database of Institute Collection and Analysis of Nanobodies (iCAN). It is built for the aim that addressing the above gap to expand and accelerate the nanobody research. iCAN, as the first database of nanobody, contains the most comprehensive information to date on nanobodies and related antigens. So far, iCAN incorporates 2391 entries which include 2131 from patents and 260 from publications and provides a simple user interface for researchers to retrieve and view the detailed information of nanobodies. In addition to the data collection, iCAN also provides online bioinformatic tools for sequence analysis and characteristic feature extraction. CONCLUSIONS: In summary, iCAN enables researchers to analyze nanobody features and explore the applications of nanobodies more efficiently. iCAN is freely available at http://ican.ils.seu.edu.cn .