Levels of free radicals and SOD in the serum in different groups of dogs.

<p>*: compared with γ-radiation alone, P<0.05.</p

Hemoglobin in Beagle dogs in different groups.

<p>Note: Data are mean +/− standard deviation. The numbers are g/L.</p><p>*: compared with model control, P<0.05;</p>#<p>: compared with positive control, P<0.05. D1-D27 are the days of the experiment.</p

Thrombocytes in Beagle dogs in different groups.

<p>Note: Data are mean +/− standard deviation. The numbers are ×10⁹/L.</p><p>*: compared with model control, P<0.05;</p>#<p>: compared with positive control, P<0.05. D1-D27 are the days of the experiment. After D7, PLT of all irradiated dogs were significantly lower than normal control, <i>P</i><0.05. Duration of thrombocytopenia is defined as the days of platelet count (PLT) is lower than 1×10¹⁰/L. Nadir (x10⁹/L) mean the lowest Thrombocytes. The recovery level (%) mean the ratio of the Thrombocytes on D27 to preradiation.</p

Activity (%) in the bone marrow of Beagle dogs in different groups.

<p>*: compared with model control, <i>P</i><0.05;</p>#<p>: compared with positive control, <i>P</i><0.05. Myelocytes∶Neutrophilic myelocytes; Metamyelocytes∶Neutrophilic Metamyelocytes; Band form: Band form neutrophils; Segmented form: Segmented form neutrophils. Basophilics∶Basophilic normoblast; Polychromatophilics∶Polychromatophilic Normoblasts; Orthochromatic∶Orthochromatic Normoblasts.</p

White blood cells in Beagle dogs in different groups.

<p>Note: Data are mean +/− standard deviation. The numbers are x10⁹/.</p><p>*: compared with model control, P<0.05;</p>#<p>: compared with positive control, P<0.05. D1-D27 are the days of the experiment. After D3, WBC of all irradiated dogs were significantly lower than normal control, P<0.05. Nadir (x109/L) mean the lowest WBC. The recovery level (%) mean the ratio of the WBC on D27 to preradiation. Duration of leukopenia is defined as the days of white blood cell count (WBC) is lower than 4x10⁹/L.</p

Hyperplasia grades in the bone marrow in different groups of dogs.

<p>*: compared with γ-radiation alone, P<0.05. Standards and scores used to grade the activity in the bone marrow are as follows. Grades include five stages of extremely active, apparently active, active, inactive, and extremely inactive. Accordingly the ratio of mature erythrocytes to karyocytes is 1∶1, 10∶1, 20∶1, 50∶1, and 300∶1 and the score is 5, 4, 3, 2, and 1 respectively.</p

Bone marrow hyperplasia of Beagle dogs irradiated by 2.0⁶°Co γ ray, at Day 28 after radiation.

<p>A. Normal control (30^#): Hyperplasia of karyocyte was apparently active. B. Model control (24^#): Hyperplasia of karyocyte was inactive. C. Positive control (16^#): Hyperplasia of karyocyte was active. D. Low dose of SNP (14^#): Hyperplasia of karyocyte was apparently active. E. Moderate dose of SNP (8^#): Hyperplasia of karyocyte was apparently active. F. High dose of SNP (1^#): Hyperplasia of karyocyte was apparently active. (Giemsa ×200).</p

DataSheet1_The pharmacological mechanism of chaihu-jia-longgu-muli-tang for treating depression: integrated meta-analysis and network pharmacology analysis.docx

Aim: Chaihu-jia-Longgu-Muli-tang (CLM) is derived from “Shang Han Lun” and is traditionally prescribed for treating depression. However, there is still a lack of evidence for its antidepressant effects, and the underlying mechanism is also unclear. This study aimed to assess clinical evidence on the efficacy of CLM in patients with depression using a meta-analysis and to explore its underlying antidepressant molecular mechanisms via network pharmacology.Methods: Eight open databases were searched for randomized controlled trials (RCTs) comparing the effects of CLM alone or combined with serotonin-norepinephrine reuptake inhibitors (SNRIs) and selective serotonin reuptake inhibitors (SSRIs) in patients with depression, evaluating the total effective rate of the treatment group (CLM alone or combined with SSRIs/SNRIs) and the control group (SNRIs or SSRIs), and comparing changes in depression scale, anxiety scale, sleep scale, inflammation indicators and adverse effects. Subsequently, the active ingredients and target genes of CLM were screened through six databases. Then Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and protein-protein interaction (PPI) network and topology analysis were performed. Finally, Molecular docking was applied to evaluate the binding affinity between components and predicted targets.Results: Twenty-four RCTs with a total of 2,382 patients were included. For the efficacy of antidepression and adverse effects, whether CLM alone or in combination with SSRIs/SNRIs, the treatment group has no inferior to that of the control group. Additionally, the intervention of CLM + SSRI significantly improved the symptoms of anxiety and insomnia, and reduced serum IL-6 and TNF-α levels. For network pharmacology, a total of 129 compounds and 416 intersection targets in CLM were retrieved. The interaction pathway between CLM and depression is mainly enriched in PI3K-Akt, JAK-STAT, and NF-κB signaling pathway, PIK3R1, MAPK3, and AKT1 may be the potential targets of Stigmasterol, β-stiosterol, coumestrol.Conclusion: Compared to SSRIs/SNRIs alone, CLM is more effective and safe in treating depression. It not only significantly alleviates depressive mood, but improves symptoms such as anxiety and insomnia, with fewer side effects, especially in combination with SSRI. Its antidepressant mechanism may be correlated with the regulation of the PI3K/Akt signaling pathway and inhibiting inflammatory response.</p

Neutrophils in Beagle dogs in different groups.

<p>Note: Data are mean +/− standard deviation. The numbers are ×10⁹/L.</p><p>*: compared with model control, P<0.05;</p>#<p>: compared with positive control, P<0.05. D1-D27 are the days of the experiment. After D3, NEUT of all irradiated dogs were significantly lower than normal control, <i>P</i><0.05. Duration of neutropenia is defined as the days of absolute neutrophil count (ANC) is lower than 2×10⁹/L. Nadir (x10⁹/L) mean the lowest Neutrophils. The recovery level (%) mean the ratio of the Neutrophils on D27 to preradiation.</p

Depletion of RAD18 suppressed entry into the M phase from the G2 phase after IR exposure.

<p>(A) HT1080 cells transfected with si-ctrl or si-RAD18 were exposed to 1 or 2 Gy IR, and then lysed at the indicated time points, after irradiation. Samples were analyzed by western blotting with the indicated antibodies. (B) Cells were exposed to 1 Gy of IR, fixed with ethanol at the indicated time points after irradiation, and then immunostained with phospho-histone H3 and propidium iodide (PI). The percentage of G2/M phase cells was determined by flow cytometry. Each value represents the mean (+standard deviation) of the results from three independent experiments. (C) Cells were exposed to various doses of IR and then fixed with ethanol 60 min after irradiation. The fixed cells were immunostained with phosphor-histone H3 and PI. The percentage of G2/M phase cells was determined by flow cytometry. Each value represents the mean (+standard deviation) of the results from three independent experiments.</p