Objective To investigate the role and underlying mechanism of C1q-like protein 4 (C1ql4) in regulating the characteristics of breast cancer stem cells. Methods qRT-PCR was used to detect the expression of C1ql4 in breast cancer and normal breast epithelial cell lines, as well as to verify the transfection efficiency of C1ql4. Western blot analysis was employed to examine the phosphorylation levels of AKT, IKK, and IκB in different groups. An AKT activator was added to MDA-MB-231 cells with C1ql4 knockdown, whereas inhibitors targeting AKT, IKK, IκB, and NF-κB nuclear translocation were separately introduced to C1ql4-overexpressing MCF-7 cells. The nuclear translocation of NF-κB, expression levels of the target genes TNF-α and IL-1β, formation ability of tumorspheres, and proportion of CD44⁺/CD24^−/low stem-like subgroups were analyzed. Results C1ql4 expression in breast cancer cell lines was significantly upregulated compared with that in normal breast epithelial cells. Western blot analysis showed that p-AKT/AKT, p-IKK/IKK, and p-IκB/IκB ratios markedly reduced in C1ql4-knockdown MDA-MB-231 cells (all P<0.05) but significantly increased in C1ql4-overexpressing MCF-7 cells (all P<0.05). Rescue experiments demonstrated that the addition of an AKT activator to C1ql4-knockdown MDA-MB-231 cells resulted in the enhanced nuclear translocation of NF-κB, the increased nuclear/cytoplasmic NF-κB ratios, the elevated TNF-α and IL-1β expression levels, and significant recovery of tumorsphere formation ability and the proportion of CD44⁺/CD24^−/low stem-like subpopulations (all P<0.05). Conversely, in C1ql4-overexpressing MCF-7 cells, treatment with AKT, IKK, IκB, or NF-κB nuclear translocation inhibitors led to a reduction in NF-κB nuclear translocation, decreased nuclear/cytoplasmic NF-κB ratios, and declines in TNF-α and IL-1β expression levels, tumorsphere formation ability, and the CD44⁺/CD24^−/low subpopulation (all P<0.05). Conclusion C1ql4 promotes the translocation of NF-κB from the cytoplasm to the nucleus through the PI3K/AKT/NF-κB signaling pathway and enhances the expression of stemness in breast cancer cells.

This study optimizes the extraction process and explores the antioxidant activity of Gastrodia elata polysaccharides, aiming to provide theoretical reference for the extraction, development, and application of the polysaccharides. Polysaccharides were extracted from G. elata by an ultrasonic-assisted method with deep eutectic solvents. The extraction process was optimized by single factor and response surface tests. The antioxidant activity of polysaccharides was evaluated by DPPH and ABTS⁺ free radical scavenging rates. The optimal deep eutectic solvents were composed of choline chloride and lactic acid at a molar ratio of 1:2. The optimal extraction conditions were the ultrasonic treatment at 50 ℃ for 48 min, a solid-to-liquid ratio of 1:38, and a water content of 42%. Under these conditions, the polysaccharide yield reached (19.88±0.93)%. The results of antioxidant activity experiment in vitro showed that the scavenging rates of G. elata polysaccharides on DPPH and ABTS⁺ free radicals were up to (26.39±1.47)% and (30.61±0.16)%, respectively, which indicated that the polysaccharides extracted by the deep eutectic solvents had a certain antioxidant ability. The extracted polysaccharides can be further studied and developed as a potential natural antioxidant.
Polysaccharides/pharmacology* ; Gastrodia/chemistry* ; Antioxidants/pharmacology* ; Deep Eutectic Solvents/chemistry* ; Solvents/chemistry*

One-day-old AA broilers were divided into five groups(15 chickens each,5 replicates per group):control(basic diet),three groups with low,medium,and high doses of crude extract of Flos sophorae and Fructus sophorae(100,150,200 mg/kg),and one group with Macleaya cordata extract(300 mg/kg).The 42-day trial measured intestinal enzyme activity,morphology,antioxidant and immune capacity,barrier function,and microbiota structure and diversity.Compared to the control and Macleaya cordata groups,the high-dose crude extract of Flos sophorae and Fructus sophorae group significantly increased trypsin activity in the duodenum,jejunum,and ileum(P＜0.05).It also reduced reactive oxygen species and malondialdehyde levels,increased glu-tathione peroxidase activity,reduced tumor necrosis factor-α,increased interleukin-10,and elevated mRNA expression of tight junction protein-1 and mucin-2 in the jejunum(P＜0.05).Microbial di-versity analysis showed higher Shannon index,increased Firmicutes and Bacteroidetes,decreased Proteobacteria,and more beneficial bacteria in the high-dose group(P＜0.05).Supplementing 200 mg/kg of crude extract of Flos sophorae and Fructus sophorae enhances intestinal morpholo-gy and function,and promotes intestinal health,thereby increasing farming efficiency.

In order to investigate the effects of Fuzi Lizhong Oral liquid on immune function and in-testinal health of chicks,185 1-day-old healthy chicks were randomly divided into 4 groups.The blank control group(CG group)was given normal tap water;the high,medium and low dose groups(FZH group,FZM group and FZL group)were given tap water containing 5.00,2.50 and 1.25 g/L of Fuzi Lizhong Oral liquid,respectively.Starting from the first day of age,the drug was administered continuously for 5 d,and the blood was collected from the subwing vein on the sixth day of the test.The results showed that compared with CG group,thymus index in FZH group was significantly increased(P＜0.05).Compared with CG group,serum IgM,IgG and sIgA in FZH group were significantly increased(P＜0.05),and serum IgG and sIgA in FZM group were signifi-cantly increased(P＜0.05).Compared with CG group,the relative expression of Occludin and Claudin-1 mRNA in FZH group was significantly increased(P＜0.05),and the relative expression of ZO-1 mRNA in FZM and FZH groups was significantly increased(P＜0.05).Compared with CG group,Shannon index of FZH group was significantly increased(P＜0.05),Simpson index of FZH group was significantly decreased(P＜0.05),and many beneficial bacteria such as Strepto-coccus spinosus,Eubacillus spinosus and Lactobacillus spinosus played a synergistic role.The results showed that adding 5.00 g/L Fuzi Lizhong Oral liquid in drinking water could improve the immunity of chicks,maintain the intestinal barrier function of chicks,increase the intestinal flora richness and promote the intestinal health of chicks.

A total of 300 1-day-old broilers were randomly divided into 5 groups with 5 replicates per group and 12 broilers per replicate.The control group was given free drinking water(CG),the astragalus polysaccharide control group(HPS)received 0.8 mL/L of astragalus polysaccharide o-ral liquid in drinking water,and the experimental groups(SBL,SBM,SBH)received 1.5,3.0,4.5 mL/,of Shenling Baizhu Oral Liquid in drinking water.The results showed as follows:com-pared to the CG group,SIgA content in HPS group,group SBM and group SBH was significantly increased(P＜0.05),IL-6 and IL-1β contents were significantly decreased(P＜0.05),Occludin,Mucin-2 and Bcl-2 contents were significantly increased(P＜0.05).The results of 16S rRNA test showed that the specific OUT number in groups HPS and SBM was significantly higher than that in group CG(P＜0.05),α diversity analysis showed that compared with group CG,Chao1 index and Simpson index of group HPS,group SBM and group SBH were significantly increased,and βdiversity analysis showed that there were significant differences in species composition between test group and blank control group(P＜0.05).The relative abundance analysis at the phylum level showed that the relative abundance of Firmicutes and Bacteroides in groups SBM and SBH was significantly higher than that in group CG(P＜0.05).The above results showed that Shenling Baizhu Oral Liquid could improve the intestinal health and enhance the resistance of broilers.

One-day-old AA broilers were divided into five groups(15 chickens each,5 replicates per group):control(basic diet),three groups with low,medium,and high doses of crude extract of Flos sophorae and Fructus sophorae(100,150,200 mg/kg),and one group with Macleaya cordata extract(300 mg/kg).The 42-day trial measured intestinal enzyme activity,morphology,antioxidant and immune capacity,barrier function,and microbiota structure and diversity.Compared to the control and Macleaya cordata groups,the high-dose crude extract of Flos sophorae and Fructus sophorae group significantly increased trypsin activity in the duodenum,jejunum,and ileum(P＜0.05).It also reduced reactive oxygen species and malondialdehyde levels,increased glu-tathione peroxidase activity,reduced tumor necrosis factor-α,increased interleukin-10,and elevated mRNA expression of tight junction protein-1 and mucin-2 in the jejunum(P＜0.05).Microbial di-versity analysis showed higher Shannon index,increased Firmicutes and Bacteroidetes,decreased Proteobacteria,and more beneficial bacteria in the high-dose group(P＜0.05).Supplementing 200 mg/kg of crude extract of Flos sophorae and Fructus sophorae enhances intestinal morpholo-gy and function,and promotes intestinal health,thereby increasing farming efficiency.

In order to investigate the effects of Fuzi Lizhong Oral liquid on immune function and in-testinal health of chicks,185 1-day-old healthy chicks were randomly divided into 4 groups.The blank control group(CG group)was given normal tap water;the high,medium and low dose groups(FZH group,FZM group and FZL group)were given tap water containing 5.00,2.50 and 1.25 g/L of Fuzi Lizhong Oral liquid,respectively.Starting from the first day of age,the drug was administered continuously for 5 d,and the blood was collected from the subwing vein on the sixth day of the test.The results showed that compared with CG group,thymus index in FZH group was significantly increased(P＜0.05).Compared with CG group,serum IgM,IgG and sIgA in FZH group were significantly increased(P＜0.05),and serum IgG and sIgA in FZM group were signifi-cantly increased(P＜0.05).Compared with CG group,the relative expression of Occludin and Claudin-1 mRNA in FZH group was significantly increased(P＜0.05),and the relative expression of ZO-1 mRNA in FZM and FZH groups was significantly increased(P＜0.05).Compared with CG group,Shannon index of FZH group was significantly increased(P＜0.05),Simpson index of FZH group was significantly decreased(P＜0.05),and many beneficial bacteria such as Strepto-coccus spinosus,Eubacillus spinosus and Lactobacillus spinosus played a synergistic role.The results showed that adding 5.00 g/L Fuzi Lizhong Oral liquid in drinking water could improve the immunity of chicks,maintain the intestinal barrier function of chicks,increase the intestinal flora richness and promote the intestinal health of chicks.

A total of 300 1-day-old broilers were randomly divided into 5 groups with 5 replicates per group and 12 broilers per replicate.The control group was given free drinking water(CG),the astragalus polysaccharide control group(HPS)received 0.8 mL/L of astragalus polysaccharide o-ral liquid in drinking water,and the experimental groups(SBL,SBM,SBH)received 1.5,3.0,4.5 mL/,of Shenling Baizhu Oral Liquid in drinking water.The results showed as follows:com-pared to the CG group,SIgA content in HPS group,group SBM and group SBH was significantly increased(P＜0.05),IL-6 and IL-1β contents were significantly decreased(P＜0.05),Occludin,Mucin-2 and Bcl-2 contents were significantly increased(P＜0.05).The results of 16S rRNA test showed that the specific OUT number in groups HPS and SBM was significantly higher than that in group CG(P＜0.05),α diversity analysis showed that compared with group CG,Chao1 index and Simpson index of group HPS,group SBM and group SBH were significantly increased,and βdiversity analysis showed that there were significant differences in species composition between test group and blank control group(P＜0.05).The relative abundance analysis at the phylum level showed that the relative abundance of Firmicutes and Bacteroides in groups SBM and SBH was significantly higher than that in group CG(P＜0.05).The above results showed that Shenling Baizhu Oral Liquid could improve the intestinal health and enhance the resistance of broilers.

Objective To explore the effects and mechanism of triptolide on proliferation and apoptosis of breast cancer MCF-7 cells.Methods MCF-7 cells were treated by different concentrations of triptolide.CCK-8 as-say was employed to detect the cell proliferation. The morphological changes were observed by an inverted micro-scope.The apoptosis rate was detected by flow cytometry.Expressions of Bcl-2,Bax,Survivin and Caspase-3 were measured by qRT-PCR and Western blot. Results Triptolide inhibited the proliferation of MCF-7 cells in a dose and time-dependent manner at a suitable range.Triptolide induced morphological changes and apoptosis.Triptolide also down-regulated Bcl-2 and Survivin expressions and up-regulated Bax and Caspase-3 expressions. Conclu-sions Triptolide inhibits proliferation and induces apoptosis of MCF-7 cells,and its mechanism may be related to down-regulation of Bcl-2 and Survivin expressions and up-regulation of Bax and Caspase-3 expressions.