Under the guidance of condensed matter theory, calcium salt cross-linking is used to prepare moisture-resistant intermediates of Yuzhu (Polygonatum odoratum) polysaccharide, reduce its hygroscopicity, improve moisture resistance and provide new methods for moisture-proofing of traditional Chinese medicine extracts.

Single-factor experiment investigated the effects of cross-linking agent type and dosage, polysaccharide concentration, stirring time, stirring speed and stirring temperature on moisture absorption rate of moisture-resistant intermediates, and used the response surface method to optimize the prescription and process. The equilibrium moisture absorption rate and critical relative humidity of moisture-proof intermediates were measured, and the glass transition temperature T_g was measured by differential scanning calorimeter (DSC), the microscopic morphology was observed by scanning electron microscope (SEM), and the rheometer was used to measure the apparent viscosity and viscoelasticity.

The type and dosage of cross-linking agent, polysaccharide concentration and stirring temperature all had significant effects on the moisture absorption rate of the intermediate. The optimal prescription and process were 5.0% calcium hydrogen phosphate, polysaccharide concentration 0.6 g/mL and stirring temperature 50 ℃. The evaluation results of moisture-resistant intermediates showed that the equilibrium moisture absorption rates of polysaccharides, polysaccharides+CaHPO₄ and moisture-resistant intermediates were 22.87%, 18.44% and 15.04%, respectively, with critical relative humidity of 51.97%, 56.53% and 66.32%, respectively, and the values of T_g were 63.5 ℃, 68.5 ℃ and 80.0 ℃, respectively. SEM showed that moisture-resistant intermediates of Polygonatum odoratum polysaccharide had a large particle size and adhesions between the particles. The rheological analysis has found that the apparent viscosity of moisture-resistant intermediates of Yuzhu (Polygonatum odoratum) polysaccharide and the elasticity increased.

Traditional Chinese medicine extracts with polymers as the active ingredient can change their condensed state structure through cross-linking, thereby reducing hygroscopicity and improving moisture resistance.

This study is based on intestinal flora-host metabolism and deeply explores the mechanism of Shanzhuyu (Corni Fructus) polysaccharides in type 2 diabetes.

A type 2 diabetes model was established by high sugar and high fat feed combined with a small dose of intraperitoneal injection of streptozocin (STZ). The pharmacoefficiency index of the rats in each group was detected, and the intestinal microbial structure of the rats in each group was detected by 16S rDNA sequencing technology, and the content of short-chain fatty acids (SCFAs) was measured in combination with GC-MS technology. The Western Blot method was used to detect the protein levels of G-protein coupled receptor 43 (GPR43), G-protein coupled receptor 41 (GPR41) and glucagon-like peptide-1 (GLP-1) in liver tissues. Small molecule metabolomics study was performed on animal feces in each group using UPLC-MS/MS technology, and the correlation analysis was conducted on intestinal microbiota and differential metabolites.

The results of the drug efficacy index test showed that Shanzhuyu (Corni Fructus) polysaccharides could significantly reduce the blood sugar, blood lipids and inflammatory factors in the rats with type 2 diabetes. The results of intestinal flora sequencing showed that Shanzhuyu (Corni Fructus) polysaccharides significantly improved the bacterial diversity of type 2 diabetes rats. At the level of phylum, the abundance of Firmicutes, Bacteroides, Varves and Actinomycetes all had different degrees of correction, and the ratio of Firmicutes/Bacteroides increased significantly (P<0.05). At the level of genus, Lactobacillus, Akermans, Bacteroides and Muribaculaceae increased. The results of SCFAs and key metabolic pathways showed that the content of SCFAs in the feces of type 2 diabetes rats had significantly decreased (P<0.01), the protein expressions of GPR43, GPR41 and GLP-1 were significantly reduced (P<0.01), and after the intervention of Shanzhuyu (Corni Fructus) polysaccharides, the SCFAs content was significantly increased (P<0.05, P<0.01), and the protein expressions of GPR41, GPR43 and GLP-1 were significantly increased (P<0.01). Metabolomics screened differential metabolites with significant changes (P<0.05) among 46 model and control groups, mainly enriched 19 metabolic pathways such as sphingolipid metabolism and steroid biosynthesis. The results of the joint analysis showed that intestinal flora had a significant regulatory effect on differential metabolites such as sphingolipid metabolism and steroid biosynthesis, involving metabolic pathways such as sphingolipid metabolism and steroid biosynthesis.

Shanzhuyu (Corni Fructus) polysaccharides can affect the levels of metabolites such as SCFAs and sphingosine by regulating the intestinal microbiota structure, and then regulate the metabolism pathways such as short-chain fatty acid metabolism, sphingolipid metabolism and steroid biosynthesis, improve the protein expressions of GPR43, GPR41 and GLP-1, regulate the blood sugar and blood lipid levels of the rats, thereby regulating glycolipid metabolism and intestinal disorders, inhibiting the body's inflammatory response, reducing insulin resistance and playing a role in treating type 2 diabetes.

To investigate the effects of different concentrations of Yinyanghuo (Epimedii Folium) extract on osteogenic differentiation of the rats adipose-derived stem cells (ADSCs).

Five healthy, nulliparous SPF-grade female SD rats were used to isolate and culture the ADSCs. The obtained third-generation ADSCs were stained with Alizarin Red and Oil Red O to evaluate their osteogenic and adipogenic differentiation abilities after being cultured with 1, 10, 50, 100 μmol/L Yinyanghuo (Epimedii Folium) extract. Then, the third-generation ADSCs were divided into a blank group, a resveratrol group (100 μmol/L) and Yinyanghuo (Epimedii Folium) extract groups. According to the different concentrations of Yinyanghuo (Epimedii Folium) extract, four subgroups were created, namely Ⅰ～Ⅳ groups (1, 10, 50, 100 μmol/L). After 7 days of culture, qRT-PCR and Western blotting were performed to detect the expression levels of LIM domain-containing protein Ajuba (Ajuba), Runx-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) in the ADSCs.

The resveratrol group and Yinyanghuo (Epimedii Folium) extract I to IV groups all upregulated the expression levels of Runx2 gene and protein, as well as ALP gene, with the best effect observed in the 100 μmol/L Yinyanghuo (Epimedii Folium) extract group (P<0.01). As for the expression levels of ALP protein, the resveratrol group exhibited a stronger promoting effect than the Yinyanghuo (Epimedii Folium) extract I to IV groups (P<0.01). The resveratrol group and Yinyanghuo (Epimedii Folium) extract groups I to IV all downregulated the expression levels of Ajuba gene and protein (P<0.01, P<0.05), with the resveratrol group showing a stronger gene inhibitory effect at the same concentration.

Both Yinyanghuo (Epimedii Folium) extract and resveratrol can promote the osteogenic differentiation of adipose-derived stem cells, but the promoting effect of Yinyanghuo (Epimedii Folium) extract is more significant and effective. Both can also inhibit adipogenic differentiation, with Yinyanghuo (Epimedii Folium) extract exhibiting weaker inhibition than resveratrol in the short term. With the extension of intervention time, further research is needed to investigate the trend in the future.

To investigate the effect and mechanism of water extract of Buyang Huanwu Decoction (补阳还五汤, BHD) on H₂O₂-induced apoptosis in PC12 cells based on the JNK/c-Jun signaling pathway.

An oxidative stress cell model was established by treating PC12 cells with H₂O₂ at an optimal concentration of 200 μmol/L. The experiment was divided into a normal group, a model group, low-, middle-and high-dose BHD groups, and an SP600125 group. PC12 cells were treated with SP600125 and different concentrations of BHD medium. The cell proliferation was detected using a real-time cell analysis (RTCA) system and the CCK-8 method. Intracellular reactive oxygen species (ROS) levels and apoptosis were measured by immunofluorescence and flow cytometry. The protein expression levels of Bax, Caspase-3, Bcl-2, c-Jun, p-c-Jun, p-JNK and JNK were detected by Western blotting.

Compared with the normal group, the model group showed significantly increased ROS levels, apoptosis rate, protein expressions of Bax and Caspase-3 and ratios of p-JNK/JNK and p-c-Jun/c-Jun, while the expression of Bcl-2 protein was significantly decreased (P<0.01). Compared with the model group, high-dose BHD intervention significantly reduced ROS levels, apoptosis rate, protein expressions of Bax and Caspase-3 and ratios of p-JNK/JNK and p-c-Jun/c-Jun (P<0.01), while significantly increasing the expression of Bcl-2 protein (P<0.01).

The water extract of Buyang Huanwu Decoction can inhibit oxidative stress-mediated apoptosis in PC12 cells through the JNK/c-Jun signaling pathway, revealing a potential mechanism by which Buyang Huanwu Decoction may improve refractory erectile dysfunction by promoting nerve injury repair.

Based on the theory of the “gut-osteoporosis” axis and 16S rDNA technology, to explore the effect and mechanism of Modified Huoluo Xiaoling Dan (加味活络效灵丹) on the repair of bone tissue in the rats with steroid-induced avascular necrosis of the femoral head (SANFH), as well as its influence on the structure of the gut microbiota.

Using the random number table method, 24 SPF-grade male SD rats were divided into a normal group (n=8), a model group (n=8) and a treatment group (n=8). In the model group and the treatment group, the method of alternately injecting methylprednisolone sodium succinate into the gluteal muscles was used to establish the model, and the treatment group was given intragastric administration of Modified Huoluo Xiaoling Dan (preparation liquid). For the femoral samples of the rats, HE staining was performed and the empty bone lacuna rate was calculated. Three-dimensional reconstruction by Micro-CT was used to analyze and measure the bone histological parameters such as the bone volume fraction (BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) and trabecular separation (Tb. Sp) in the measurement area. Immunohistochemistry method was used to detect the expressions of bone tissue repair markers BMP, OPG and Runx2. For the fecal samples, 16S rDNA technology was used for gut microbiota sequencing and ASV analysis. The α-diversity and β-diversity were used to analyze the species diversity of the gut microbiota, and LEfSe analysis was used to detect the changes in the species differences of the gut microbiota at the family level and genus level.

Modified Huoluo Xiaoling Dan could significantly improve the empty bone lacuna rate of the femoral head (P<0.05) and bone histological parameters such as BV/TV, Tb. N, Tb. Th and Tb. Sp (P<0.01) in the rats with SANFH, and significantly up-regulate the expression levels of bone tissue repair markers BMP, OPG and Runx2 (P<0.01). The results of gut microbiota sequencing showed that α-diversity analysis indicated that there were obvious differences in the species diversity and richness of the gut microbiota structures of the rats in the three groups; β-diversity analysis showed that the flora structure of the treatment group was significantly separated from that of the model group, and was similar to that of the blank group. The difference among groups was greater than that within groups, and the difference was obvious. LEfSe analysis has found that at the family level and genus level, the gut microbiota of the rats in the model group showed significant “pro-inflammatory” characteristics, while the gut microbiota of the rats in the treatment group showed “anti-inflammatory-promoting repair” characteristics.

The effect of Modified Huoluo Xiaoling Dan on the repair of bone tissue in the rats with SANFH is closely related to its regulation of the gut microbiota. The intervention drug effectively reshapes the intestinal microecology disturbed by hormones, further confirming that the reshaping of the flora has a bidirectional regulatory effect on the “gut-osteoporosis” axis, playing a role in preventing and treating SANFH.