Salvia miltiorrhiza ameliorates endometritis in dairy cows by relieving inflammation, energy deficiency and blood stasis.

Introduction: According to traditional Chinese veterinary medicine, endometritis is caused by a combination of Qi deficiency, blood stasis, and external evil invasion. Salvia miltiorrhiza is a traditional Chinese medicine that counteracts blood stasis and has additional demonstrated effects in boosting energy and restraining inflammation. Salvia miltiorrhiza has been employed in many traditional Chinese prescriptions that have proven effective in healing clinical dairy cow endometritis. Methods: the in vivo effect of Salvia miltiorrhiza in treating endometritis was evaluated in dairy cows. In addition, bovine endometrial epithelium cell inflammation and rat blood stasis models were employed to demonstrate the crosstalk between energy, blood circulation and inflammation. Network analysis, western blotting, qRT-PCR and ELISA were performed to investigate the molecular mechanism of Salvia miltiorrhiza in endometritis treatment. Results: The results demonstrate that treatment with Salvia miltiorrhiza relieves uterine inflammation, increases blood ATP concentrations, and prolongs blood clotting times. Four of the six Salvia miltiorrhiza main components (SMMCs) (tanshinone IIA, cryptotanshinone, salvianolic acid A and salvianolic acid B) were effective in reversing decreased ATP and increased IL-1ß, IL-6, and IL-8 levels in an in vitro endometritis model, indicating their abilities to ameliorate the negative energy balance and external evil invasion effects of endometritis. Furthermore, in a blood stasis rat model, inflammatory responses were induced in the absence of external infection; and all six SMMCs inhibited thrombin-induced platelet aggregation. Network analysis of SMMC targets predicted that Salvia miltiorrhiza may mediate anti-inflammation via the Toll-like receptor signaling pathway; anti-aggregation via the Platelet activation pathway; and energy balance via the Thermogenesis and AMPK signaling pathways. Multiple molecular targets within these pathways were verified to be inhibited by SMMCs, including P38/ERK-AP1, a key molecular signal that may mediate the crosstalk between inflammation, energy deficiency and blood stasis. Conclusion: These results provide mechanistic understanding of the therapeutic effect of Salvia miltiorrhiza for endometritis achieved through Qi deficiency, blood stasis, and external evil invasion.

Effect of different acupuncture sequences of Huiyangjiuzhen acupoints on blood glucose and hemorheology in the anesthetized rabbits.

Background and objective: Hemorheology and blood glucose are commonly used to estimate the risks of thrombosis and stress hyperglycemia after anaesthesia. The sequence of acupoint stimulation might influence the therapeutic effects of acupuncture. In the current study, we aimed at investigating the effect of different acupuncture sequences of "Huiyangjiuzhen" acupoints on the blood glucose and hemorheology in anesthetized rabbits. Methods: Twenty-five rabbits were randomly divided into five groups, including the control group (CG), the positive-sequence group (PSG), the reverse-sequence group (RSG), the disorder-sequence group (DSG), and the random group (RG). Except for the CG and RG, the rabbits in other groups were acupunctured with different sequences of "Huiyangjiuzhen"acupoints when the rabbits were anesthetized. The acupoints in rabbits of the RG were chosen randomly. The levels of blood glucose and hemorheology indexes before and after anaesthesia was detected. Results: In the PSG, Hηb 200/s, Mηb 30/s, Hηr 200/s, ERI, hematocrit and plasma viscosity levels were decreased, and the blood glucose level was not changed. In the DSG, the levels of Mηb 30/s and hematocrit were decreased, and the blood glucose was increased. In the CG, RSG and RG, no hemorheology indexes were changed and the blood glucose was increased. Conclusion: "Huiyangjiuzhen" acupuncture could decrease the risks of post-operative thrombosis and stress hyperglycemia in anesthetized rabbits. This effectiveness depends on both acupuncture and acupuncture sequence at the "Huiyangjiuzhen" acupoints.

Four Patterns of Canine Wei Syndrome Treated with Traditional Chinese Medicine.

Atrophy and weakness of the limbs is a common condition in animals, especially dogs. It typically presents with flaccidity and weakness of the limbs, especially the hind legs, muscle atrophy, and the inability to walk. In Traditional Chinese Medicine (TCM) and Traditional Chinese Veterinary Medicine (TCVM), this is known as wei syndrome (WS). According to TCM, the etiology of WS can be (1) lung heat and fluid consumption; (2) insufficiency of the liver and kidneys; (3) dampness-heat invasion; (4) damage to the spleen and stomach, which are also the patterns of WS. This report aims to provide an alternative option for the treatment of canine paralysis. Four dogs with different WS patterns were treated with acupuncture, moxibustion, and Chinese herbs based on the guidelines of the TCM literature. Three patients recovered normal functioning. The fourth patient could walk normally after 2 weeks of treatment, but his hind limbs became weak again 3 months later. Weekly acupuncture treatment was resumed until his death 18 months later. TCM application of acupuncture, moxibustion, and Chinese herbs can be an effective treatment for canine WS. It is hoped that this case report will broaden the treatment options of other veterinarians when patients present with this condition.

Research progress on the extraction technology and activity study of Epimedium polysaccharides.

More pharmacological effects of polysaccharides from traditional Chinese medicines have been discovered in recent years. Epimedium has been used for thousands of years as a traditional Chinese medicine in China. Water-soluble Epimedium polysaccharides is one of the main ingredients of Epimedium, which is one of the main active ingredients of Epimedium, mainly composed of mannose, rhamnose, galacturonic acid, glucose, and galactose. The extraction methods of Epimedium polysaccharides including hot water extraction, cellulase extraction, ultrasonic extraction, microwave-assisted extraction, ultrasound compound enzyme and ultra-high pressure extraction, they affect the yield of Epimedium polysaccharides. The characteristics of deproteinization including enzyme deproteinization, macroporous resin deproteinization and Sevag methods are introduced respectively. Some chemical modification methods of Epimedium polysaccharides are also involved such as phosphorylation, sulfation, selenization, and lipids encapsulated. Epimedium polysaccharides have a variety of pharmacological activities, including immune promotion, reproduction promotion, anti-osteoporosis, anti-tumor, antioxidant, anti-fatigue and antivirus, also beneficial to nervous and hematopoietic systems. At present, the research of Epimedium polysaccharides has been in depth. In this paper, the research progress on extraction, purification, chemical modification methods and pharmacological activity of Epimedium polysaccharides summarized. The aim is to provide reference for further research and development of Epimedium polysaccharides.

Study on the SHP2-Mediated Mechanism of Promoting Spermatogenesis Induced by Active Compounds of Eucommiae Folium in Mice.

Spermatogenesis directly determines the reproductive capacity of male animals. With the development of society, the increasing pressure on people's lives and changes in the living environment, male fertility is declining. The leaf of Eucommia ulmoides Oliv. (Eucommiae Folium, EF) was recorded in the 2020 Chinese Pharmacopoeia and was used in traditional Chinese medicine as a tonic. In recent years, EF has been reported to improve spermatogenesis, but the mechanisms of EF remain was poorly characterized. In this study, the effect of EF ethanol extract (EFEE) on spermatogenesis was tested in mice. Chemical components related to spermatogenesis in EF were predicted by network pharmacology. The biological activity of the predicted chemical components was measured by the proliferation of C18-4 spermatogonial stem cells (SSCs) and the testosterone secretion of TM3 leydig cells. The biological activity of chlorogenic acid (CGA), the active compound in EF, was tested in vivo. The cell cycle was analysed by flow cytometry. Testosterone secretion was detected by ELISA. RNA interference (RNAi) was used to detect the effect of key genes on cell biological activity. Western blotting, qRT-PCR and immunofluorescence staining were used to analyse the molecular mechanism of related biological activities. The results showed that EFEE and CGA could improve spermatogenesis in mice. Furthermore, the main mechanism was that CGA promoted SSC proliferation, self-renewal and Leydig cell testosterone secretion by promoting the expression of SHP2 and activating the downstream signaling pathways involved in these biological processes. This study provided strong evidence for elucidating the mechanism by which EF promotes the spermatogenesis in mice and a new theoretical basis for dealing with the decrease in male reproductive capacity.

Extraction, purification, structural character and biological properties of propolis flavonoids: A review.

Propolis is an aromatic substance which is collected by bees and mixed with bee saliva. The plant sources of propolis are mainly consisted with plant exudates from bark, buds and etc. Flavonoids are secondary metabolites widely found in natural plants, which have a variety of health care functions and are the main active ingredients of propolis. This article summarized the types, active ingredients, pharmacological effects, extraction methods and applications of propolis flavonoids, the aim was to provide the theoretical basis for further research and development of propolis flavonoids.

Bioactivity-guided isolation of immunomodulatory compounds from the fruits of Ligustrum lucidum.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Ligustrum lucidum (FLL) W.T. Aiton (Oleaceae) is included in the 2020 "Chinese Pharmacopoeia" and is widely used in traditional Chinese medicine as a tonic. In recent years, FLL has been reported to improve immune function, but the bioactive compounds and mechanisms of FLL remain poorly characterized. AIM OF THE STUDY: To identify FFL compounds with strong immune activity and explore their molecular mechanisms. MATERIALS AND METHODS: The phagocytic activity of RAW264.7 macrophages and proliferation activity of spleen lymphocytes were used to guide the isolation of bioactive compounds from FLL extracts. Lymphocyte subpopulations, Ca2+ concentrations, and surface molecule expression were analyzed using flow cytometry. Cytokine secretion was examined using ELISA. FITC-OVA uptake was observed using fluorescence microscopy. NF-κB activation was analyzed using western blotting. RESULTS: The extraction and isolation produced ten compounds, namely oleuropeinic acid, nuezhenide, isonuezhenide, salidroside, isoligustrosidic acid, ligulucidumosides A, 8(E)-nuezhenide, hydroxytyrosol, oleuropein, and p-hydroxyphenethyl 7-ß-D-glucosideelenolic acid ester were isolated and identified from FLL-Bu-30%. Immunoactivity experiments showed that hydroxytyrosol had the strongest macrophage phagocytotic and lymphocyte proliferation-promoting activities. Further studies showed that hydroxytyrosol could significantly enhance lymphocyte subsets CD3+, CD4+/CD8+, and CD3+CD4-CD8-, promote IL-4, IFN-γ, and TNF-α secretion, and increase intracellular Ca2+ concentrations. In addition, the results from RAW264.7 macrophages showed that hydroxytyrosol increased FITC-OVA uptake, induced TNF-α and IL-1ß production, upregulated MHC-II, CD80, and CD86 expression, promoted cytoplasmic IκB-α degradation, and increased nuclear NF-κB p65 levels. CONCLUSION: Our study provides substantial evidence regarding the mechanism of the immunomodulatory effects of compounds from FLL.

Characterization and antioxidant activity of the oligo-maltose fraction from Polygonum Cillinerve.

In this study, Polygonum Cillinerve polysaccharide (PCP) was extracted, and characterized by high performance gel permeation chromatography, high performance liquid chromatography, fourier transform infrared spectroscopy, nuclear magnetic resonance and mass spectroscopy. The results showed that PCP was composed of glucose, it was α-D-glucan and the backbone of PCP was consisted of repeating units of (1→4)-α-D-Glucose. In addition, the antioxidant potential of PCP was assessed in vitro. The results showed that PCP had strong hydroxyl radical scavenging ability, some DPPH scavenging ability and good reducing power. Moreover, the results of the enzyme-linked immunosorbent assays showed that PCP at 15.625-0.975 µg/mL could significantly improve the level of superoxide dismutase and glutathione peroxidase, and could significantly decrease the level of myeloperoxidase, malondialdehyde and xanthine oxidase in macrophages. These results indicated that PCP could potentially be developed as a natural antioxidant.

The acaricidal activity and mechanism of eugenol on Psoroptes cuniculi.

In this study, the acaricidal effect of eugenol was measured and its mechanism of action investigated. The results showed that eugenol possessed the effect of killing Psoroptes cuniculi, and could regulate the mRNA expression of glutathione S-transferase (GST), catechinic acid (Ca) and thioredoxin (Trx). PPAR, NF-kappa B, TNF, Rap 1 and Ras signaling pathways might be the main pathways that involved into the process of killing mites. These findings suggested that eugenol could be developed into a new kind of acaricide, and further expand current knowledge on the mechanisms of eugenol for killing Psoroptes cuniculi of eugenol.

Phospholipase Cγ2 signalling contributes to the haemostatic effect of Notoginsenoside Ft1.

OBJECTIVES: The drawback of bleeding caused by chronic antiplatelet therapy is persecuting patients with thrombotic diseases severely. Based on the dual-directional regulatory effect of Panax notoginseng on platelet, the present study focused on the effect of Notoginsenoside Ft1, a saponin with effect in promoting platelet aggregation. KEY FINDINGS: According to the present study, Notoginsenoside Ft1 cannot stimulate platelet aggregation independently. However, the effect in enhancing aggregation induced by thrombin, collagen and ADP is peaked at 5-10 µm. In addition, thrombin-induced activation of PLCγ2-IP3 /DAG-[Ca2+ ]/PKC-TXA2 signalling was potentiated by Notoginsenoside Ft1, as well. Furthermore, the mice tail bleeding time was shortened by administration of Notoginsenoside Ft1 significantly. And the bleeding time prolonged by aspirin was also restored by Ft1. CONCLUSIONS: The haemostatic effect of Notoginsenoside Ft1 was exerted through potentiation of PLCγ2-IP3 /DAG-[Ca2+ ]/PKC-TXA2 signalling pathway stimulated by other stimulators. Notoginsenoside Ft1 has the potential to be developed into supplements in antiplatelet therapy to prevent the drawback of bleeding.

Coptis chinensis and Myrobalan (Terminalia chebula) Can Synergistically Inhibit Inflammatory Response In Vitro and In Vivo.

Objectives. To investigate the anti-inflammatory effect of Coptis chinensis plus myrobalan (CM) in vitro and in vivo. Methods. The inflammation in mouse peritoneal macrophages was induced by lipopolysaccharide (LPS). Animal models were established by using ear swelling and paw edema of mouse induced by xylene and formaldehyde, respectively. In vitro, cytotoxicity, the phagocytosis of macrophages, the levels of nitric oxide (NO), induced nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in cell supernatant were detected. In vivo, swelling rate and edema inhibitory rate of ear and paw were observed using CM-treated mice. Results. At 150-18.75 µg·mL(-1), CM had no cytotoxicity and could significantly promote the growth and the phagocytosis of macrophages and inhibit the overproduction of NO, iNOS, TNF-α, and IL-6 in macrophages induced by LPS. In vivo, pretreatment with CM, the ear swelling, and paw edema of mice could be significantly inhibited in a dose-dependent manner, and the antiedema effect of CM at high dose was better than dexamethasone. Conclusion. Our results demonstrated that Coptis chinensis and myrobalan possessed synergistically anti-inflammatory activities in vitro and in vivo, which indicated that CM had therapeutic potential for the prevention and treatment of inflammation-mediated diseases.