The purpose of this paper is to explore the decoction and dosing methods of rhubarb root and rhizome in classical prescriptions and to provide a reference basis for the clinical use of rhubarb root and rhizome. By collating the relevant classical prescriptions of rhubarb root and rhizome in Shanghan Lun and Jingui Yaolüe, the relationship between its decoction and dosing methods and the syndrome was analyzed. The decoction of rhubarb root and rhizome in classical prescriptions can be divided into three categories: simultaneous decoction, decoction later, and other methods (impregnation in Mafei decoction, decoction with water from the well spring first taken in the morning, and pills). If it enters the blood level or wants to slow down, rhubarb root and rhizome should be decocted at the same time with other drugs. If it enters the qi level and wants to speed up, rhubarb root and rhizome should be decocted later. If it wants to upwardly move, rhubarb root and rhizome should be immersed in Mafei decoction. If it wants to suppress liver yang, rhubarb root and rhizome should be decocted with water from the well spring first taken in the morning. If the disease is prolonged, rhubarb root and rhizome should be taken in pill form. The dosing methods of rhubarb root and rhizome can be divided into five categories: draught, twice, three times, before meals, and unspecified. For acute and serious illnesses with excess of pathogenic qi and adequate vital qi, we choose draught. For gastrointestinal diseases, we choose to take the medicine twice. For achieving a moderate and long-lasting effect, we choose to take the medicine three times. If the disease is located in the lower part of the heart and abdomen, we choose to take it before meals. The use of rhubarb root and rhizome in clinical practice requires the selection of the appropriate decoction and dosing methods according to the location of the disease, the severity of the disease, the patient′s constitution, and the condition after taking the medicine.

BACKGROUND:During healing process of chronic wounds,excessive production of reactive oxygen species can impair the function of L929 fibroblasts,thereby delaying wound repair.Therefore,protecting fibroblasts from oxidative stress is important to promote wound healing. OBJECTIVE:To assess the protective effects of carboxymethyl chitosan-oxidized chondroitin sulfate/platelet-rich plasma(CMC-OCS/PRP)hydrogel on L929 cells under H2O2 stimulation. METHODS:CMC-OCS/PRP hydrogels were prepared,and the micromorphology,degradation performance,scavenging ability of H2O2 and hydroxyl radical and biocompatibility of the hydrogels were characterized.L929 cells with good growth state were taken and cultured in five groups.The control group was cultured conventionally.H2O2 was added to the H2O2 group.Carboxymethyl chitosan-oxidized chondroitin sulfate hydrogel extract+H2O2 was added to the CMC-OCS group.Platelet-rich plasma gel extract+H2O2 was added to the PRP group.The CMC-OCS/PRP group was treated with carboxymethyl chitosan-oxidized chondroitin sulfate/platelet-rich plasma hydrogel extract+H2O2.Each group was treated with hydrogel extract for 6 hours,and then H2O2 for 24 hours.After culture,the levels of active oxygen and malondialdehyde,apoptosis and expression of collagen fiber I protein were detected.In the presence of H2O2,the above hydrogel extracts were directly or indirectly co-cultured with L929 fibroblasts for 36 hours,respectively.Migration ability of the cells was detected by scratch test and Transwell chamber test. RESULTS AND CONCLUSION:(1)CMC-OCS/PRP hydrogels had uniform and interrelated porous structure and good degradation ability,could effectively remove H2O2 and hydroxyl radicals in vitro,and had good biocompatibility.(2)Compared with the control group,the apoptosis rate,reactive oxygen species,and malondialdehyde levels were increased(P<0.05);the spread area of cells was decreased(P<0.05),and the expression of collagen fiber I protein had no significant changes(P>0.05)in the H2O2 group.Compared with the H2O2 group,reactive oxygen species level was decreased in the CMC-OCS group(P<0.05),malondialdehyde level was decreased(P<0.05),and cell spread area was increased(P<0.05)in the PRP group,CMC-OCS group,and CMC-OCS/PRP group;apoptosis rate was decreased in the CMC-OCS/PRP group(P<0.05),and collagen fiber I protein expression was increased in the PRP group,CMC-OCS group,and CMC-OCS/PRP group(P<0.05).(3)Compared with the control group,the number of cell migration was decreased(P<0.05),and the migration area had no significant change(P>0.05)in the H2O2 group.Compared with the H2O2 group,the number and area of cell migration were increased in the PRP group,CMC-OCS group,and CMC-OCS/PRP group(P<0.05),and the increase was most significant in the CMC-OCS/PRP group.(4)Under oxidative stress,CMC-OCS/PRP hydrogel can improve the migration ability of fibroblasts,resist cell apoptosis,and preserve cell extension function.

" Lijie and yellowish sweating" originates from the chapter on stroke and arthralgia diseases in Synopsis of Golden Chamber. Later generations typically interpret it as yellow fluid oozing from painful joints, a characteristic manifestation of arthralgia. In Western medicine, Lijie corresponds to diseases such as gouty arthritis, with its primary clinical manifestations being redness, swelling, heat, and painful joints, most often without yellow fluid discharge. Therefore, the interpretation of " Lijie and yellowish sweating" contradicts the clinical manifestations often observed in this disease. Thus, this article reinterprets the meaning of " Lijie and yellowish sweating" from the pathogenesis of " sweat exposure to water, as if water harms the heart" , combined with the viewpoints of other medical practitioners. Determining the meaning of " yellowish sweating" is crucial for understanding the pathogenesis of arthralgia and clarifying the relationship between arthralgia and yellowish sweating. ZHANG Zhongjing mentioned arthralgia and " yellowish sweating" together, not to differentiate between the two diseases but to emphasize the common pathogenesis of the two, namely, the cold and dampness injuring the heart, blood, and vessels. This paper proposes a new explanation of " Lijie and yellowish sweating" , which suggests that " yellowish sweating" is not confined to the joints but can be found all over the body. The pathogenesis of " Lijie and yellowish sweating" lies in the insufficiency of the liver and kidney and exogenous water dampness, leading to disharmony between nutrient qi and defensive qi and between yin and yang. Primary treatment should harmonize yingfen and weifen, as well as tonify and replenish the liver and kidney. The clinical selection of medicines can be considered Guizhi Decotion, a type of formula. The pathogenesis of " Lijie and yellowish sweating" is complex, and clinical treatment should be comprehensively considered to achieve the best therapeutic effect.

Autism spectrum disorder in children (hereinafter called "childhood autism") is a prevalent neurodevelopmental condition; however, ancient Chinese medical texts did not provide detailed accounts of it. The concept of "the sovereign fire illuminates, while the ministerial fire occupies its position" was first introduced in the Plain Questions: The Great Treatise on the Origin of Heavenly Influences chapter. Subsequent medical practitioners have elaborated on this idea, identifying the sovereign fire as the master of the spirit and governing life activities, while the ministerial fire is the assisting minister that supports the functions of the zang-fu organs. When these two are in harmony, they complement each other, collectively sustaining the body′s normal physiological functions. In the context of this theory, the pathogenesis of childhood autism is attributed to the imbalance between the sovereign and ministerial fires. The root cause lies in the mutual antagonism of these two fires, resulting in the dysfunction of the spirit. Central to childhood autism onset is the dimming of the sovereign fire and a deficiency in the spirit, while the displacement of the ministerial fire and the impaired function of the five viscera contribute significantly to the progression of the condition. Effective treatment for childhood autism involves restoring these imbalances: dispelling yin-related obstructions to illuminate the sovereign fire, nourishing the spleen and kidney to nourish the ministerial fire, and regulating the liver and lung to harmonize the sovereign and ministerial fires. The ultimate goal is to integrate body and spirit, harmonize the sovereign and ministerial fires, and restore the interconnectedness between spirit and the function of the five zang viscera. This article, based on the "the sovereign fire illuminates, while the ministerial fire occupies its position" theory, provides a novel perspective and insights for understanding and treating childhood autism through the holistic principles of traditional Chinese medicine. Emphasizing the integration of body and spirit offers new insights and approaches for the clinical management of childhood autism in traditional Chinese medicine.

Probiotics have shown excellent application prospects in preventing and treating many diseases. However, their sensitivity to the harsh environment in vivo always leads to a massive loss of viability and insufficient therapeutic effect. Fortunately, modified probiotics have emerged and provide multiple possibilities for their use in various diseases. Modification not only endows probiotics with extra capacity to resist severe environments but also gives them exogenous characteristics, such as prolonged retention time and improved therapeutic effects. Modified probiotics could combine with other therapies, which has opened up new avenues to enhance the efficacy of probiotic-based therapy. In this review, we have summarized the current physicochemical and biological modification strategies of probiotics. In addition, the progress of research on probiotic-based combination therapy has also been extensively reviewed, which contributes to the enhanced delivery of probiotics or other active constituents and provides new ideas for disease treatment, bioimaging, and diagnosis.

Psoriasis is a chronic inflammatory skin disease, and its pathogenesis is largely modulated by abnormal angiogenesis. Previous research has indicated that AlkB homolog 5 (ALKBH5), an important demethylase affecting N⁶-methyladenosine (m⁶A) modification, plays a role in regulating angiogenesis in cardiovascular and eye diseases. Our present study found that ALKBH5 was upregulated and co-localized with cluster of differentiation 31 (CD31) in the skin of IMQ group compared with control group. ALKBH5-deficient mice decreased IMQ-induced psoriatic dermatitis and exhibited histological improvements, including decreased epidermal thickness, hyperkeratosis, numbers of dermal capillary vessels and inflammatory cell infiltration. ALKBH5-KO mice alleviated angiogenesis in psoriatic lesions by downregulating the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. Additionally, the expression of ALKBH5 was significantly upregulated in IL-17A-induced human umbilical vein endothelial cells (HUVECs), which further promoted the expression of angiogenesis-related cytokines and endothelial cell proliferation. Cell proliferation and angiogenesis were suppressed in ALKBH5 knockdown group, whereas ALKBH5 overexpression promoted these processes. The regulation of angiogenesis in HUVECs by ALKBH5 was facilitated through the AKT-mTOR pathway. Collectively, ALKBH5 plays a pivotal role in psoriatic dermatitis and angiogenesis, which may offer a new potential targets for treating psoriasis.
Animals ; Psoriasis/chemically induced* ; Mice ; Humans ; Neovascularization, Pathologic/genetics* ; Human Umbilical Vein Endothelial Cells/metabolism* ; AlkB Homolog 5, RNA Demethylase/genetics* ; Proto-Oncogene Proteins c-akt/metabolism* ; TOR Serine-Threonine Kinases/metabolism* ; Cell Proliferation ; Mice, Knockout ; Disease Models, Animal ; Signal Transduction ; Male ; Skin/blood supply* ; Mice, Inbred C57BL ; Angiogenesis

The culture of suspended Vero cells is facing difficulties such as low cell viability and long doubling time. To investigate the main reasons for the slow growth and low viability of suspended Vero cells, this study conducted transcriptomic analysis of suspended Vero cells (Vero-XF) and adherent Vero cells (Vero-AD) to screen the differentially expressed genes (DEGs) affecting the growth of suspended cells. In addition, epidermal growth factor (EGF) was supplemented to the culture system to improve the growth of Vero-XF. The results showed that compared with the Vero-AD group, the Vero-XF group had 7 376 significant DEGs. Kyoto encyclopedia of genes and genomes enrichment analysis revealed that the DEGs were mainly enriched in the autophagy and mitophagy pathways. Eleven DEGs were selected and verified by quantitative real-time PCR, which showed up-regulated expression of ATG9B, WIPI2, LAMP2, OPTN, Rab7a, and DEPTOR and down-regulated expression of ATG4D, being consistent with the results of transcriptomic analysis. In addition, the Vero-XF group showed significantly up-regulated expression of ATG101, ATG2A, and STX17 and insignificant change in the expression of NBR1, compared with the Vero-AD group. The protein levels of LC3 and P62 in Vero-XF and Vero-AD were determined by Western blotting, which showed up-regulated expression of LC3Ⅱ/Ⅰ and down-regulated expression of P62 in Vero-XF, indicating a higher level of autophagy. Finally, the exogenous supplementation of EGF at 10, 20, and 30 μg/L in the culture system reduced the autophagy level of Vero-XF by 22.35%, 48.15%, and 71.29%, increased the specific growth rate by 15.48%, 33.33%, and 57.14%, and decreased the apoptosis rate by 2.84%, 15.46%, and 16.23%, respectively. The results of this study preliminarily reveal that the activation of autophagy is one of the reasons for the slow growth of Vero-XF, which provides reference for the subsequent culture of suspended Vero cells.
Animals ; Vero Cells ; Autophagy/genetics* ; Chlorocebus aethiops ; Epidermal Growth Factor/pharmacology* ; Gene Expression Profiling ; Transcriptome ; Cell Survival

ObjectiveTo explore the antioxidant and anti-human cervical cancer HeLa cell effect and mechanisms of Andrographis paniculata polysaccharide （APP）. MethodCell function assays were conducted to assess the effects of APP （400， 450， 500 mg·L^-1） on the proliferation， migration， and invasion of HeLa cells using the cell counting kit-8 （CCK-8） assay， scratch assay， and Transwell assay. Molecular mechanism experiments were conducted to detect the effects of APP on HeLa cell apoptosis and cell cycle-related mRNA and protein expression using flow cytometry， real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）， and Western blot analysis. The antioxidant activity of APP was tested using DPPH⁺， OH^-， and reducing power assays. ResultCompared with the blank group， APP （400， 450， 500 mg·L^-1） significantly inhibited the migration， proliferation， and invasion abilities of HeLa cells in a time- and dose-dependent manner （P<0.05， P<0.01）. Flow cytometry with propidium iodide （PI） single staining was used to detect cell cycle. The results showed that compared with the blank group， the proportion of HeLa cells in the G₂/M phase increased after 48 hours of treatment with APP （400， 450， 500 mg·L^-1）， indicating that APP can arrest HeLa cells in the G₂/M phase. Flow cytometry with fluorescein isothiocyanate （Annexin V-FITC）/PI apoptosis kit was used to detect cell apoptosis. Compared with the blank group， the proportion of early and late apoptotic HeLa cells increased in a dose-dependent manner after 48 hours of APP （400， 450， 500 mg·L^-1） treatment （P<0.05， P<0.01）， indicating that APP promotes HeLa cell apoptosis. The results of Real-time PCR and Western blot assay showed that compared with the blank group， after 48 hours of APP （400， 450， 500 mg·L^-1） treatment resulted in decreased mRNA and protein expression of cell cycle-dependent kinase-1 （CDK-1）， Cyclin B₁， and B-cell lymphoma-2 （Bcl-2）， and increased mRNA and protein expression of cysteine aspartate protease （Caspase）-3， Caspase-8， Caspase-9， and Bcl-2-associated X protein （Bax） （P<0.05， P<0.01）. These findings were consistent with the flow cytometry results and showed a dose-dependent effect. In vitro antioxidant tests demonstrated that different concentrations of APP （50-1 000 mg·L^-1） were able to scavenge DPPH⁺ and OH^- radicals， indicating certain antioxidant activity. ConclusionAPP possesses antioxidant activity and can inhibit the viability of HeLa cells while promoting their apoptosis.

ObjectiveTo explore the research status and trends of cohort studies on traditional Chinese medicine （TCM） efficacy evaluation from 2017 to 2022 and provide ideas and references for research in this field. MethodsSix databases including Pubmed， Web of Science， Embase， Scopus， Cochrane Library and CNKI were searched from January 1st， 2017 to December 31st， 2022. The total number of annual publications， journals， highly cited literatures， and keywords were quantitatively and visually analyzed by Bibliometrix. ResultsA total of 328 articles were included， which were published in 141 journals. The number of articles published in this field showed an overall upward trend， and retrospective cohort studies （282 papers， 85.98%） accounted for the largest proportion. A total of 151 cohort studies （46.04%） were conducted based on the database and showed an overall upward trend. The subjects were mainly patients with tumors （77 papers， 23.48%）， and cardiovascular and cerebrovascular diseases （64 papers， 19.51%）. The top 3 highly cited literatures mainly explore the association between TCM and survival outcome and quality of life in patients with malignant tumors. Fourteen and twenty-five high-frequency keywords were included in Chinese and English literature respectively， which formed 3 clusters such as research methods， statistical analysis and diseases. ConclusionIt was the current status to focus on retrospective cohort studies and focus on patients with tumors or cardiovascular and cerebrovascular diseases. Using observational database to conduct cohort studies of TCM efficacy evaluation could be the future research direction.

BACKGROUND:Diabetic wounds have complicated conditions such as infection,ischemia,peripheral neuropathy,and vascular disease.Ordinary hydrogel dressings with single structure and function cannot meet the needs of diabetic wound healing. OBJECTIVE:To explore the effect of a hydrogel loaded with platelet-rich plasma on wound healing of full-thickness skin defects in diabetic rats. METHODS:The blood of SD rats was extracted to prepare platelet-rich plasma.Carboxymethyl chitosan/oxychondroitin sulfate hydrogel and carboxymethyl chitosan/oxychondroitin sulfate hydrogel loaded with platelet-rich plasma were prepared separately.Streptozotocin was used to induce diabetes model in adult male SD rats.A round full-thickness skin wound with a diameter of 2 cm was made on the back of diabetic rats.The rats were randomly divided into four groups(n=10 per group).The blank group was applied with gauze on the wound.The hydrogel group,platelet-rich plasma group,and composite hydrogel group were respectively applied with the corresponding hydrogel,platelet-rich plasma and hydrogel loaded with platelet-rich plasma.The wound healing was observed within 20 days after treatment. RESULTS AND CONCLUSION:(1)On day 20 after treatment,the wound healing rate of the hydrogel group,platelet-rich plasma group and composite hydrogel group was significantly higher than that of the blank group(P<0.05).The wound healing rate of the composite hydrogel group was significantly higher than that of the platelet-rich plasma group(P<0.05).(2)The results of hematoxylin-eosin staining on day 5 after treatment showed that compared with the blank group,hydrogel group and platelet-rich plasma group,there were a large number of inflammatory cell infiltration,new granulation tissue and capillary formation in the wound tissue of the composite hydrogel group.(3)On day 5 after treatment,the results of immunohistochemical staining and western blot assay showed that the expression levels of tumor necrosis factor α and interleukin 1β in wound tissue in the composite hydrogel group were significantly lower than those in the blank group,hydrogel group and platelet-rich plasma group(P<0.05).(4)Masson staining results on day 15 after treatment showed that compared with the blank group,hydrogel group and platelet-rich plasma group,there were more collagen fibers in the wound tissue of the composite hydrogel group,which were orderly,evenly distributed and dense.(5)CD31 immunofluorescence staining showed that on day 15 after treatment,the expression of CD31 in wound tissue of the composite hydrogel group was significantly higher than that of the blank group,hydrogel group and platelet-rich plasma group(P<0.05).(6)These results suggest that the hydrogel loaded with platelet-rich plasma can promote the healing of full-thickness skin defect wounds in diabetic rats by promoting granulation tissue,collagen fiber and angiogenesis,and reducing the inflammatory response.