Slow transit constipation (STC), the most common type of constipation, seriously affects the life of patients. Zhizhu decoction (ZZD), a traditional Chinese medicine compound, has is effective ...against functional constipation, but the mechanism is still unclear.
This research explores the mechanism of ZZD on STC from the perspective of metabolomics and gut microbiota.
Fifty-four C57BL/6 mice were randomly divided into six groups (n = 9): control (control); STC (model); positive control (positive); low-dose (5 g/kg; L-ZZD), medium-dose (10 g/kg; M-ZZD), and high-dose (20 g/kg; H-ZZD) ZZD treatment. Following treatment of mice with ZZD for two weeks, the changes in intestinal motility, colon histology, intestinal neurotransmitters, and aryl hydrocarbon receptor (AHR) pathway determined the effects of ZZD on the pathophysiology of STC. LC-MS targeting serum metabolomics was used to analyze the regulation of ZZD on neurotransmitters, and 16S rRNA high-throughput sequencing was used to detect the regulation of the gut microbiome.
ZZD had the highest content of naringin (6348.1 mg/L), and could significantly increase the 24 h defecations (1.10- to 1.42-fold), fecal moisture (1.14-fold) and intestinal transport rate (1.28-fold) of STC mice, increased the thickness of the mucosal and muscular tissue (1.18- to 2.16-fold) and regulated the neurotransmitters in the colon of STC mice. Moreover, ZZD significantly activated the AHR signaling pathway, and also affected the composition of gut microbiota in STC mice.
The beneficial effect and the possible mechanism of ZZD on STC could provide a theoretical basis for the broader clinical application of ZZD.
Colorectal cancer is one of the most common cancers, and its incidence rate keeps increasing worldwide. Epidermal growth factor receptor (EGFR) is highly expressed in colorectal cancer cells, and its ...activation leads to the activation of downstream kinase pathways to promote cell growth, resulting in tumour growth and progression. EGFR is expressed in only small amounts in normal cells, however, it is overexpressed in tumour cells and is a potential tumour marker, thus the detection of which is of great importance. This work proposes a novel and highly sensitive electrochemical immunosensor based on nucleic acid aptamer recognition and silver deposition for EGFR detection. The key novelty of this work is the use of SiO2 nanospheres to enhance the sensitivity. First, SiO2 nanospheres were synthesized and modified on the surface of a gold electrode using a self-assembly process. This provided a large surface area to allow high loading of capture antibodies. The antibody was then immobilized on the SiO2 nanosphere modified electrode surface to form an immunosandwich complex with the antigen and biotin-labelled nucleic acid aptamer. Afterwards, the affinity-labelled alkaline phosphatase was immobilized onto the electrode with the specific reaction of biotin and affinity. The obtained product was subjected to silver deposition in the substrate solution. The large surface area provided by the SiO2 nanospheres allowed significantly improved sensitivity. Experiments were performed to optimize the concentration of immobilized antibodies and biotin-labelled nucleic acid aptamer. The experimental results show a good linearity of EGFR concentration in the range of 1 to 1000 ng/mL, with the lowest detection limit up to 0.06 ng/mL. This method has high sensitivity and stability, which allows it to have a broad application prospects in the clinical field.
Bian-Se-Tong mixture (BSTM) is an optimized formulation based on the classical prescription “Zhizhu pill”, which is widely used in the clinical treatment of slow-transit constipation (STC). The ...potential molecular mechanism of BSTM therapy for STC was investigated by network pharmacology prediction combined with animal experiments. The active components of BSTM were screened via the TCMSP platform. The GeneCards, OMIM and DrugBank databases were used to search for STC targets. With the help of the Biogenet tool, a protein interaction network between drugs and disease targets was constructed, and the intersection network of the two was extracted to obtain the key targets of BSTM in the treatment of STC. GO and KEGG enrichment analyses of key targets were carried out with Metascape. Loperamide hydrochloride was used to establish an STC rat model, and the key targets and related pathways were preliminarily verified. The important signaling pathways included the PI3K-Akt, MAPK, IL-17, cAMP, and cell cycle signaling pathways. The experimental results showed that BSTM treatment increased the body weight of STC rats and increased the fecal particle number, fecal water content and intestinal carbon ink promotion rate within 24 h. Further pathological changes in the colon of the rats were also observed. In-depth mechanistic studies have shown that BSTM can significantly reduce the apoptosis of intestinal Cajal cells, downregulate the expression of Bax and c-Caspase 3, upregulate the expression of Bcl-2 and c-kit, and promote the phosphorylation of AKT. The results showed that BSTM can significantly relieve constipation in STC rats via a mechanism related to activating the PI3K-Akt signaling pathway and improving Cajal cell apoptosis.
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•This study reported for the first time that Bian-Se-Tong mixture could significantly improve the apoptosis of Cajal cells in STC model rats.•This study reported for the first time that Bian-Se-Tong mixture could significantly improve the pathological status of STC model rats through the method of network pharmacology combined with experimental verification.•This study reported that Bian-Se-Tong mixture may protect Cajal cells damage in STC model rats by regulating PI3K/AKT signaling pathway.
Postoperative wound of perianal infectious disease represents a common but unique refractory wound in clinical practice. The reasons that hinder the wound healing process include not only the severe ...bacterial infection of the wound itself and the narrow and deep shape of the wound, but also its frequent bacterial contact. Therefore, the development of biofunctional dressings to aid in therapy is essential. In this study, we synthesized a new type of dressing comprising a hydrogel host based on the Schiff base principle and catechol groups between polydopamine, oxidized dextran, and quaternized chitosan, and then loaded it with traditional Chinese medicine molecules. These formed an integrated hydrogel for accelerated wound repair in a perianal infection model. The prepared hydrogels exhibited excellent wet tissue adhesion, antifouling, morphological variability, suitable swelling properties, and complete degradability, as well as remarkable contact antibacterial ability and the ability to rapidly scavenge free radicals. Hemostatic experiments showed excellent hemostatic properties, as the integrated hydrogel could instantly gel to seal the hemorrhage. Hemocompatibility and
in vitro
cell experiments showed that the integrated hydrogel had good biosafety and significantly promoted cell proliferation, which in turn accelerated the repair of infected whole cortexes in rats. A histomorphological evaluation showed that the integrated hydrogel promoted the recovery of normal anatomical tissue in rats by promoting the formation of collagen fibers and inhibiting inflammation. The results showed that this multifunctional integrated hydrogel has great potential for the treatment of continuously infected skin regeneration, providing a promising therapeutic strategy for postoperative wound healing in perianal infectious diseases.
Astragaloside IV (AS‐IV) has exhibited pivotal anti‐cancer efficacy in multiple types of cancer, including colorectal cancer (CRC). Meanwhile, circular RNA (circRNA) circ_0001615 has been reported to ...be involved in the malignant development of CRC. Herein, this study is expected to figure out the interaction between circ_0001615 and AS‐IV on CRC progression. The 50% inhibition concentration (IC50), proliferation, apoptosis, and migration were detected by Cell Counting Kit‐8 (CCK‐8), 5‐ethynyl‐2′‐deoxyuridine (EdU), flow cytometry, and wound healing assays. The expression of related proteins was examined by western blot. Circ_0001615, microRNA‐873‐5p (miR‐873‐5p), and LIM and SH3 protein 1 (LASP1) levels were detected by real‐time quantitative polymerase chain reaction (RT‐qPCR). The binding between miR‐873‐5p and circ_0001615, or LASP1, was predicted by Starbase, followed by verification by dual‐luciferase reporter and RNA immunoprecipitation (RIP) assays. The biological role of circ_0001615 and AS‐IV on CRC tumor growth was detected by the xenograft tumor model in vivo. According to the IC50 of AS‐IV in CRC cells, the 100 ng/mL AS‐IV treatment for 24 h was chosen for the following research: Our data confirmed that AS‐IV is a beneficial anti‐cancer agent in CRC cells. Furthermore, circ_0001615 and LASP1 expression were increased, and miR‐873‐5p was decreased in CRC patients and cell lines, whereas their expression exhibited an opposite trend in AS‐IV‐treated cells. Functionally, applying AS‐IV might act as a beneficial anti‐cancer effect by downregulating circ_0001615 in CRC cells in vitro. Mechanically, circ_0001615 serves as a sponge for miR‐873‐5p to affect LASP1 expression. In addition, AS‐IV inhibited CRC cell growth in vivo by modulating circ_0001615. Overall, AS‐IV could mitigate CRC development, at least in part, through the circ_0001615/miR‐873‐5p/LASP1 axis. These findings support a theoretical basis for an in‐depth study of the function of AS‐IV and the clinical treatment of CRC.
Applying astragaloside IV (AS‐IV) might repress the expression of circ_0001615 in colorectal cancer (CRC) cell lines. Furthermore, AS‐IV performed a beneficial anti‐cancer effect by regulating the circ_0001615/miR‐873‐5p/LASP1 network in the CRC.
Constipation is a frequent gastrointestinal symptom. It is intimately related to many diseases. 1st-line therapy can not alleviate constipation for some patients. Alternative treatments are therefore ...commonly used, such as probiotics. Nevertheless, the efficacy and safety of probiotics used as a single treatment are still uncertain. A systematic review and meta-analysis will be carried out to answer the issue.
The protocol accompanied Preferred Reporting Items for Systematic Reviews and Protocol Meta-Analysis. PubMed, Cochrane, Embase, and Web of Science databases were practiced for randomized controlled trials without language constraint. In addition, We have also conducted backward (manually) and forward (with Google Scholar) citation checks to identify any additional relevant papers.Two reviewers will conduct studies selection, data extraction, and risk of bias assessment independently. The primary outcome is treatment success (spontaneous bowel movements (sBMs) >3 times per week), defecation frequency. The second result will be consistency, fecal incontinence, other symptoms (e.g. flatulence, abdominal pain), and adverse event rates and types.
This study provides helpful information about whether probiotics can be used as a single therapy on functional constipation CONCLUSION:: The findings of the review will be disseminated through peer-review publications.
To explore the function and regulation mechanism of circ_0071589 in colorectal cancer (CRC). The expression levels of circ_0071589, microRNA-296-5p (miR-296-5p), and Engrailed-2 (EN2) were detected ...by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot was performed to check the protein levels of EN2 and apoptosis-related proteins. Cell colony formation and 5-Ethynyl-29-deoxyuridine (EdU) assay were used to exhibit cell proliferation. Cell apoptosis was shown by flow cytometry. Tube formation assay manifested the angiogenesis ability of CRC cells. Transwell assay demonstrated cell migration and invasion. The interaction between miR-296-5p and circ_0071589 or EN2 was identified by dual-luciferase reporter assay. The effect of circ_0071589 on tumor formation was demonstrated by in vivo tumor formation experiments. Immunohistochemical (IHC) assay was used to detect the positive cell rate of Ki67 in tumor tissue. Circ_0071589 was upregulated in CRC tissue and cells. Circ_0071589 knockdown repressed CRC cells proliferation, angiogenesis, migration, invasion, and promoted cell apoptosis. MiR-296-5p was downregulated in CRC tissue and cells. And miR-296-5p inhibitor could reverse the malignant phenotypes and angiogenesis inhibition of CRC cells caused by circ_0071589 knockdown. Additionally, miR-296-5p decreased CRC cell colony formation, EdU-positive cells, angiogenesis, and increased cell apoptosis through reducing the expression level of EN2. Finally, circ_0071589 silencing inhibited tumor formation in vivo. Circ_0071589 upregulated EN2 expression through sponging miR-296-5p, thereby promoting the malignant phenotype and angiogenesis of CRC cells, which provided a new target for the treatment of CRC.
Background. Maren pills have been used to treat constipation. Aquaporin 3 (AQP3) plays a vital role in regulating water transfer in the colon. It has been reported that the downregulation of AQP3 can ...regulate liquid water metabolism and intestinal permeability in irritable bowel syndrome (IBS) rats’ colon via NF-κB pathway. In this study, we investigated whether the laxative effect of Maren pills is associated with the regulation of AQP3 and NF-κB signaling pathway in the colon. Methods. The compound diphenoxylate suspension-induced STC rats received Maren pills intragastrically for 1 consecutive week to evaluate the laxative effect of Maren pills involving the regulation of AQP3 and NF-κB signaling pathway. Moreover, human intestinal epithelial cells (HT-29) were treated with drug serum to obtain in vitro data. Results. Our results revealed that treatment with Maren pills increased the stool number, moisture content of feces, and intestinal transit rate in a dose-dependent manner. Maren pills significantly increased the AQP3, fibrosis transmembrane conductance regulator (CFTR), and protein kinase A (PKA) proteins in the colon of rats and in HT-29 cells. Mechanistically, Maren pills obviously inhibited the activation of NF-κB pathway in the colon of rats and in HT-29 cells. Conclusion. These results suggest that the laxative effect of Maren pills is associated with the increased expression of AQP3 by downregulating NF-κB signal pathway.
Aurantii Fructus
Immaturus
total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate ...AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network.
Zinc finger proteins (ZNFs) serve key roles in tumor formation and progression; however, the functions and underlying mechanisms of dysregulated ZNF384 in colorectal cancer (CRC) are yet to be fully ...elucidated. Therefore, the present study initially aimed to investigate the expression levels of ZNF384 in CRC samples. Moreover, lentiviral ZNF384 overexpression and ZNF384 knockdown models were established in CRC cells. Transwell, wound healing and
tail vein metastasis assays were carried out to evaluate the effects of ZNF384 on CRC cell metastasis. Furthermore, reverse transcription‑quantitative PCR, western blotting, serial deletion, site‑directed mutagenesis, dual‑luciferase reporter and chromatin immunoprecipitation assays were conducted to investigate the potential underlying mechanisms. The results of the present study demonstrated that ZNF384 expression was markedly increased in CRC samples and this was associated with a poor prognosis. Notably, ZNF384 overexpression increased the levels of CRC cell invasion and migration, whereas ZNF384 knockdown inhibited CRC development. Moreover, the results of the present study demonstrated that ZNF384 mediated the expression of MMP2. MMP2 knockdown inhibited ZNF384‑mediated CRC cell invasion and migration, whereas MMP2 overexpression ameliorated ZNF384 knockdown‑induced inhibition of CRC progression. In addition, the results of the present study demonstrated that hypoxia‑inducible factor 1α (HIF‑1α) had the ability to bind to the ZNF384 promoter, thereby initiating ZNF384 expression. In human‑derived CRC samples, the expression levels of ZNF384 were positively correlated with both MMP2 and HIF‑1α expression. Collectively, these findings highlighted that ZNF384 may act as a prognostic marker and regulator of CRC metastasis.
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