Transglutaminase cross-linking ovalbumin-flaxseed oil emulsion gels: Properties, microstructure, and performance in oxidative stability.

This study prepared emulsion gels by modifying ovalbumin (OVA)-flaxseed oil (FSO) emulsions with transglutaminase (TGase) and investigated their properties, structure and oxidative stability under different enzyme reaction times. Here, we found prolonged reaction times led to the transformation of α-helix and ß-turn into ß-sheet and random coil. The elasticity, hardness and water retention of the emulsion gels increased significantly, but the water-holding capacity decreased when the reaction time exceeded 4 h. Confocal laser scanning microscope (CLSM) indicated extended enzyme reaction time fostered oil droplet aggregation with proteins. Emulsion gel reduced FSO oxidation, especially after 4 h of the enzyme reaction, the peroxide value (PV) of the emulsion gel was reduced by 29.16% compared to the control. In summary, the enzyme reaction time of 4 h resulted in the formation of a dense gel structure and enhanced oxidative stability. This study provides the potential applications in functional foods and biomedical fields.

Porphyrin-anthracene covalent organic frameworks for sustainable photosterilization.

Covalent organic frameworks (COFs) have promising applications in enhanced phototherapy. However, COFs that can sustainably play a role in phototherapy without continuous irradiation are extremely scarce. Herein, we report the fabrication of porphyrin-anthracene multifunctional COFs (Por-DPA) for sustainable photosterilization and bacterial-infected wound healing. A porphyrin photosensitizer, as one of the monomers, was used to provide photothermal and photodynamic activities under irradiation. An anthracene derivative, a good chemical source of singlet oxygen (1O2), was selected as another monomer to capture 1O2 and release it continuously via cycloreversion in the dark. The prepared Por-DPA COF prevents the self-aggregation quenching of the photosensitizer and thermal damage caused by continuous exposure to external light sources. Besides, Por-DPA exhibits good photothermal conversion performance and efficient 1O2 production capacity through dual pathways of photosensitization and cycloreversion. The developed sustainable photosterilization platform not only has good bactericidal effects on Escherichia coli and Staphylococcus aureus, but also promotes wound healing without obvious side effects, and is expected to be a novel efficient bactericide.

Neuroprotective neolignan glycosides from the pseudobulbs of Bletilla striata.

Four undescribed neolignan glycosides, bletineosides A-D (1-4) were isolated from the pseudobulbs of Bletilla striata. Their structures with absolute configurations were elucidated on the basis of spectroscopic analyses, along with acidic hydrolysis reactions and ECD experiments. All isolates were evaluated for their neuroprotective activities against glutamate-induced PC12 cell injury. Compound 3 and 4 showed significantly neuroprotective effects at the concentration of 10 µM when compared with the model group. Compounds 1-4 represented the first examples of neolignan glycosides from the genus Bletilla. This study disclosed the potency of Bletilla striata as a new source of anti-neurodegenerative agents.

Nephrotoxicity assessment of podophyllotoxin-induced rats by regulating PI3K/Akt/mTOR-Nrf2/HO1 pathway in view of toxicological evidence chain (TEC) concept.

Adverse reactions to traditional Chinese medicine have hindered the healthy development and internationalization process of the traditional Chinese medicine industry. The critical issue that needs to be solved urgently is to evaluate the safety of traditional Chinese medicine systematically and effectively. Podophyllotoxin (PPT) is a highly active compound extracted from plants of the genus Podophyllum such as Dysosma versipellis (DV). However, its high toxicity and toxicity to multiple target organs affect the clinical application, such as the liver and kidney. Based on the concurrent effects of PPT's medicinal activity and toxicity, it would be a good example to conduct a systematic review of its safety. Therefore, this study revolves around the Toxicological Evidence Chain (TEC) concept. Based on PPT as the main toxic constituent in DV, observe the objective toxicity impairment phenotype of animals. Evaluate the serum biochemical indicators and pathological tissue sections for substantial toxic damage results. Using metabolomics, lipidomics, and network toxicology to evaluate the nephrotoxicity of PPT from multiple perspectives systematically. The results showed that PPT-induced nephrotoxicity manifested as renal tubular damage, mainly affecting metabolic pathways such as glycerophospholipid metabolism and sphingolipid metabolism. PPT inhibits the autophagy process of kidney cells through the PI3K/Akt/mTOR and Nrf2/HO1 pathways and induces the activation of oxidative stress in the body, thereby causing nephrotoxic injury. This study fully verified the feasibility of the TEC concept for the safety and toxicity evaluation of traditional Chinese medicine. Provide a research template for systematically evaluating the safety of traditional Chinese medicine.

Application and Effectiveness of Chinese Medicine in Regulating Immune Checkpoint Pathways.

Immunotherapy targeting immune checkpoint molecules has emerged as a key approach in cancer treatment, representing the forefront of antitumor research. However, studies on immune checkpoint molecules have mainly focused on targeted therapies. Chinese medicine (CM) research as a complementary medicine has revealed that immune checkpoint molecules also undergo disease-specific changes in the context of autoimmune diseases. This review article presents a comprehensive analysis of CM studies on immune checkpoint molecules in the last 5 years, with a focus on their role in different diseases and treatment modalities. CM research predominantly utilizes oral administration of herbal plant extracts or acupuncture techniques, which stimulate the immune system by activating specific acupoints through temperature and needling. In this study, we analyzed the modulation and mechanisms of immune checkpoint molecules associated with different coinhibitory and costimulatory molecules, and reviewed the immune functions of related molecules and CM studies in treating autoimmune diseases and tumors. By summarizing the characteristics and research value of CM in regulating immune checkpoint molecules, this review aims to provide a useful reference for future studies in this field.

[Effects of herbal cake separated moxibustion on Tim-4 and ubiquitination of PD-1 in rabbits with immunosuppression].

OBJECTIVE: To observe the effects of herbal cake separated moxibustion on macrophage effector molecule T-cell immunoglobulin and mucin-domain containing-4 (Tim-4) and ubiquitination of programmed cell death protein 1 (PD-1) in rabbits with immunosuppression, and to explore the possible mechanism on herbal cake separated moxibustion in improving immunosuppression. METHODS: Thirty-two big-ear white rabbits were randomly divided into a normal group, a model group, a moxa stick moxibustion group and a herbal cake separated moxibustion group, 8 rabbits in each group. Except the normal group, the immunosuppression model was established by intraperitoneal injection of cyclophosphamide of60 mg/kg in the other 3 groups. "Shenque" (CV 8), "Shenshu" (BL 23), "Zusanli" (ST 36), etc. were selected in both the moxa stick moxibustion group and the herbal cake separated moxibustion group. Moxa stick moxibustion was applied in the moxa stick moxibustion group, one cone at each acupoint; herbal cake separated moxibustion was applied in the herbal cake separated moxibustion group, 5 cones at each acupoint. The intervention was given once every other day for 10 times in both groups. Leukocyte content in peripheral blood was detected by blood cell analyzer; the positive expression of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood was measured by flow cytometry, the serum levels of interleukin 2 (IL-2), CD8, CD68 and Tim-4 were detected by ELISA, and the expression of Tim-4 and F-box only protein 38 (FBXO38) in the liver and spleen tissues was measured by immunohistochemistry. RESULTS: Compared with the normal group, in the model group, white blood cell count (WBC) and percentage of neutrophils (NEU%) were decreased while percentage of lymphocyte (LYM%) was increased (P<0.01) in peripheral blood; the positive expression rates of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood were increased (P<0.01); the serum levels of IL-2, CD68 and Tim-4 were increased (P<0.01), the serum level of CD8 was decreased (P<0.01); the average optical density (AOD) of Tim-4 in the liver tissue and FBXO38 in the liver and spleen tissues was increased (P<0.01). Compared with the model group, in the moxa stick moxibustion group and the herbal cake separated moxibustion group, WBC and NEU% were increased (P<0.01); the positive expression rates of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood were decreased (P<0.01); the serum levels of IL-2, CD68 and Tim-4 were decreased (P<0.01), the serum levels of CD8 were increased (P<0.01); the AOD of Tim-4 and FBXO38 in the liver tissue and FBXO38 in the spleen tissue was decreased (P<0.01, P<0.05). Compared with the moxa stick moxibustion group, in the herbal cake separated moxibustion group, the positive expression rate of PD-1 in CD+68 macrophages in peripheral blood was increased (P<0.05); serum level of Tim-4 was increased (P<0.01); AOD of Tim-4 in the liver tissue was decreased (P<0.05). CONCLUSION: Herbal cake separated moxibustion can improve immunosuppression by regulating the expression of macrophage effector molecule Tim-4 and the FBXO38 mediated ubiquitination of PD-1, Tim-4 may be one of the specific indexes of immunomodulation involving with herbal cake separated moxibustion.

Application of Meditation Relaxation Training and Rosenthal Effect in Patients with Adenoidectomy.

Objective: This study aims to explore the application effect of meditation relaxation training and the Rosenthal effect in patients with adenoidectomy. Methods: This study included 94 children who underwent adenoidectomy in our hospital from April 2020 to May 2022 and were divided into a study group and a control group. The control group was given routine care, and the study group was given meditation relaxation training and the Rosenthal effect on the basis of the control group. The negative emotions, treatment compliance, complication rates, and nursing satisfaction of children's family members before and after the intervention were compared between the two groups. Results: The results of this study showed that after the intervention, the CDI and SCARED scores of the children in the study group were significantly lower than those in the control group. The treatment compliance in the study group was significantly higher than that in the control group, and the incidence of complications was significantly lower than that in the control group. Conclusion: The intervention of meditation relaxation training and the Rosenthal effect on children with adenoidectomy can relieve their negative emotions, improve treatment compliance, reduce the incidence of complications, and the children's family members are more satisfied.

[Effect of herbal cake-separated moxibustion on behavioral stress reactions and blood lactic acid level and muscular AMPK/PGC-1α signaling in rats with chronic fatigue syndrome].

OBJECTIVE: To observe the effect of herbal cake-separated moxibustion (HCSM) on serum lactic acid (BLA) level and AMPK/PGC-1α signaling pathway in the quadriceps femoris in chronic fatigue syndrome (CFS) rats, so as to explore its mechanisms underlying improvement of CFS. METHODS: According to the random number table, 50 SD rats were divided into blank control, model, HCSM, sham HCSM and medication (herbal medicine gavage) groups, with 10 rats in each group. The CFS model was established by using chronic restraint and exhaustive swimming, alternately, once daily for 21 days. The herbal cake was made of Xiaoyao Powder (Mental Ease Powder, composed of [Danggui (Radix Angelicae Sinensis), Baishao (Radix Paeoniae Alba), Chaihu (Radix Bupleuri), Fuling (Poria), Baizhu (Rhizoma Atractylodis, Macrocephalae), etc.]. The HCSM was applied to "Shenque" (CV8), "Guanyuan "(CV4), bilateral "Zusanli" (ST36) and "Qimen" (LR14), 5 moxa-cones for each acupoint, once daily for 10 days. For sham HCSM, the excipient was instead of herbal cake, and the same 5 moxa-cones was given as the HCSM group. Rats of the medication group received gavage of Xiaoyao Powder suspension (60 mg·kg-1), once daily for 10 days. The open field test and tail suspension test were conducted for determining the animals' locomotor activity. The blood sample was taken from the abdominal aorta under anesthesia for assaying the levels of serum BLA, chemokine ligand CXCL9 and ß-endorphin (EP) by ELISA. Bilateral quadriceps femoris were sampled for observing histopathological changes after staining with conventional H.E. technique, and for detecting the expression levels of phosphorylated AMP-activated protein kinase (p-AMPK) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) by using immunohistochemistry. RESULTS: Compared with the blank control group, the number of rearing and horizontal grid-crossing times, struggling times of tail suspension test were significantly decreased (P<0.05), and the immobility time was obviously prolonged (P<0.05) in the model group. Compared with the model group, both HCSM and medication groups had a significant increase of rearing, horizontal grid-crossing times and struggling times (P<0.05), and the immobility time had a significant decrease (P<0.05). But there were no significant differences in the total movement distance among the 5 groups (P>0.05), and in the 5 indexes of behavioral measurements between the HCSM and medication groups (P>0.05). The sham HCSM could also evidently increase the struggling times and reduce the immobility time (P<0.05). The contents of serum BLA, CXCL9 and ß-EP were obviously higher in the model group than in the blank control group (P<0.05), as well as remarkably lower in the HCSM and medication groups than in the model group (P<0.05). Whereas the expression levels of muscular p-AMPK and PGC-1α were considerably lower in the model group than in the blank control group (P<0.05), and significantly increased in both HCSM and medication groups relevant to the model group (P<0.05). Compared with the sham HCSM group, the contents of BLA, CXCL9 and ß-EP in serum of the HCSM group and contents of CXCL9, ß-EP in medication group were significantly decreased (P<0.05), and the protein expressions of p-AMPK and PGC-1α in quadriceps femoris in both HCSM and medication groups were significantly increased (P<0.05). H.E. staining showed smaller intercellular space, uneven cytoplasmic staining in some muscle fibers, nucleus pyknosis and condensation, and inflammatory cell infiltration in the model group, which was milder in both HCSM and medication groups. CONCLUSION: HCSM can mitigate the stress behavioral state in CFS rats, which may be related with its functions in lowering the levels of serum BLA, CXCL9 and ß-EP, and activating AMPK/PGC-1α signaling pathway (balancing energy metabolism) in the quadriceps femoris.

Berberine protects against chronic cerebral hypoperfusion-induced cognitive impairment and hippocampal damage via regulation of the ERK/Nrf2 pathway.

Vascular cognitive impairment caused by chronic cerebral hypoperfusion (CCH) seriously affects the quality of life of elderly patients and places a great burden on society and family. With the development of traditional Chinese medicine (TCM), TCM approaches to the prevention and treatment of senile ischemic cerebrovascular disease has received increasing attention. In this study, rats with bilateral common carotid artery occlusion (BCCAO) were treated with berberine (BBR). Their learning and memory function, neuronal injury and repair, the extracellular regulatory protein kinase (ERK)/nuclear factor-E2-related factor 2 (Nrf2) signaling pathway, and impairment and improvement of the blood-brain barrier (BBB) were evaluated. This study found that BBR can alleviate the pathological injury to the brain, reduce neuronal loss and promote neuronal cell survival after CCH by interfering with the ERK/Nrf2 signaling pathway. BBR can reduce BBB injury in CCH rats by inhibiting the expression of VEGF-A and MMP-9 in plasma, which reveals a protective effect of BBR on vascular cognitive impairment. This study provides a new research direction for BBR in the treatment of ischemic cerebrovascular disease.

[Clinical efficacy and safety evaluation of acupuncture combined with moxibustion in treatment of allergic rhinitis].

OBJECTIVE: To evaluate the clinical efficacy and safety of acupuncture combined with moxibustion on allergic rhinitis. METHODS: Using the random number table, 80 patients with allergic rhinitis were divided into a medication group and an acupuncture combined with moxibustion (acu-mox) group, 40 cases in each one. In the medication group, ioratadine tables were prescribed for oral administration, one tablet daily for 10 days as 1 session , 3 sessions of treatment were required. In the acupuncture combined with moxibustion group, bilateral Yingxiang (LI20), Yintang (EX-HN3), bilateral Hegu (LI4) and bilateral Shenshu (BL23) were selected as the main points and stimulated with acupuncture and moxibustion; and the acupoint prescription was modified according to symptoms. This combined treatment was given once every day, stimulating for 30 min each time, and 10 treatments made 1 course, for 3 courses of treatment totally. Before and after treatment, the scores for symptoms and physical signs, as well as the score of rhino-conjunctivitis related quality of life scale (R-QOL) were evaluated separately. The sample of the inferior turbinate mucosa tissue was collected and the distribution of eosinophil (EOS) was scored using HE staining and Sheldeny evaluation. Using enzyme-linked immunosorbent assay (ELISA), the contents of serum immunoglobulin E (IgE), retinoic-acid-receptor-related orphan nuclear receptor γt (RORγt), forkhead box protein P3 (Foxp3), interleukin-17 (IL-17), IL-27 and IL-33 were determined. The clinical efficacy was evaluated in the patients with allergic rhinitis of two groups and all the adverse reactions were recorded during treatment. RESULTS: The scores of symptoms and physical signs as well as the score of R-QOL, and EOS distribution score and the contents of serum IgE, RORγt, IL-17 and IL-33 were all reduced as compared with those before treatment in each group (P<0.05), and the contents of serum Foxp3 and IL-27 were increased as compared with those before treatment in each group (P<0.05). After treatment, the scores of symptoms and physical signs as well as the score of R-QOL, and the contents of serum IgE, RORγt and IL-33 in the acu-mox group were lower than those in the medication group (P<0.05), and the contents of serum Foxp3 and IL-27 were higher than those of the medication group (P<0.05). The total effective rate of the acu-mox group was 100.0% (40/40), significantly higher than 82.5% (33/40) in the medication group (P<0.05). No ob-vious adverse reaction was found in either group during and after treatment. CONCLUSION: Acupuncture combined with moxibustion is significantly effective and safe in treatment of allergic rhinitis. Its effect mechanism may be related to the balance modulation of Th1/Th2 and Th17/Treg cells mediated by naive CD4+T cells.

Epicoccum layuense Causing Leaf Spot on Oxalis corymbosa in China.

Oxalis corymbosa DC. is a perennial herb used as the groundcover in the urban landscape and traditional Chinese medicine. In May 2019, a leaf spot-like disease was observed on about 20% of O. corymbosa plants in one park in Hefei, Anhui, China (N31°52´, E117°15´). Symptoms appeared as yellowish, irregular or circular areas that later turned pale brown, coalesced and produced large necrotic areas of various shapes. Ten leaves with typical symptoms were removed from 5 different O. corymbosa plants (2 leaves per plant) and disinfected with 75% ethanol for 30 s. The tissue pieces (4×4 mm) were cut from the margin of the leaf lesion, dipped in 2% NaClO for 1 minute, washed 3 times with sterile distilled water, placed on Potato Dextrose Agar (PDA) medium with 50 µg/ml each of ampicillin and kanamycin, and cultured at 25℃ under a 12-h light/dark cycle. The white mycelia grew out of each leaf tissue pieces 3 days later and the hyphal tips were sub-cultured into a new PDA medium for purification. After 5 days of incubation at 25℃, the produced colonies were yellow with irregular white margin from the top with floccose aerial mycelia, ranged in a diameter from 60 to 70 mm, and produced red orange pigments into PDA medium. Colonies on the reverse side were dark red. Conidia produced on PDA were ranged in size from 5.5 to 35.5 µm in diameter (n = 800), subglobose to pyriform, and yellowish-brown. The representative isolate YHF0519 was chosen for further analysis. Fragments of four genes including the internal transcribed spacer (ITS), beta-tubulin, 28S large subunit ribosomal RNA gene (LSU), and the second largest subunit of RNA polymerase II (RPB2) were amplified from the extracted genomic DNA of YHF0519 and sequenced. The corresponding primer sets were ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass and Donaldson 1995), LSU1Fd/LR5 (Crous et al. 2009; Vilgalys and Hester 1990), and RPB2-5F2/fRPB2-7cR (Sung et al. 2007; Liu et al. 1999), respectively. All resulting sequences were deposited into GenBank and received assigned accession numbers: MN418006 for ITS; MN428043 for beta-tubulin; MN428044 for LSU; MZ437946 for RPB2. A phylogenetic tree generated by the maximum likelihood method with 1,000 bootstrapping replications based on the combined ITS, beta-tubulin, LSU, and RPB2 sequences revealed that YHF0519 was clustered closest to the isolate CGMCC 3.18362 of Epicoccum layuense Qian Chen, Crous & L. Cai (Chen et al. 2017; Jayasiri et al. 2017). The pathogenicity test was performed on leaves of healthy O. corymbosa plants in the field and repeated twice. The average air temperature was about 28°C during the pathogenicity test. A total of 126 and 127 leaves were used for treatment and control, individually. Leaves were inoculated by spraying 30 ml of spore suspension (1×106 spores/ml) of YHF0519 or sterilized distilled water. Eight day-post inoculation, about 92% of the leaves inoculated with spores developed pale brown spots, which are identical to those occurred naturally in the field. No symptoms were detected on control leaves. The fungus was re-isolated from the symptomatic leaf tissues and showed the same morphological and molecular characteristics as YHF0519. E. layuense has been reported on Perilla sp., oat, and tea plant (Chen et al. 2017; Chen et al. 2019; Chen et al. 2020). This is the first report of E. layuense causing leaf spot on O. corymbosa in China. Due to the potential threat on the ornamental and medicinal value of O. corymbosa by E. layuesne, it will be necessary to develop local management strategies against leaf spot on O. corymbosa.

[Mechanism of hepatotoxicity induced by ethanol extract of Dysosma versipellis based on "quantity-weight-evidence" network toxicology].

In this study, the toxicological/pharmacological research method of "quantity-weight-evidence" network was first proposed and practiced to supplement the existing methodology of network toxicology. We transformed the traditional qualitative network into a quantitative network in this study by attributing weights to toxic component content and target frequency, which improved the reliability of data and provided a research idea for the systematic safety evaluation and toxicological research of Chinese medicinal herbs. Firstly, 50% ethanol extract of Dysosma versipellis(DV) was administrated to rats via gavage and the potential hepatotoxic components were identified by serum pharmacochemistry. Then, the component targets were obtained from SwissTargetPrediction, PharmMapper and other online databases, and the target weights were given according to the relative content of components and target fishing frequency. Meanwhile, the targets of hepatotoxicity were predicted from online databases such as Comparative Toxicology Database(CTD) and GeneCards. Subsequently, protein-protein interaction analysis and KEGG pathway enrichment were performed with the STRING database. Finally, the quantitative network of "toxic components-weighted targets-pathways" was constructed. Eleven potential toxic compounds were predicted, including podophyllotoxin, podophyllotoxone, deoxypodophyllotoxin, and 6-methoxypodophyllotoxin. A total of 106 hepatotoxic targets and 65 weighted targets(e.g., Cdk2, Egfr, and Cyp2 c9) were identified. The results of Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment showed that these targets could act on PI3 K-AKT, MAPK, and Ras signaling pathways to play a role in inflammatory response and oxidative stress. However, traditional network toxicology showed that 51 targets such as AKT1, Alb, and Stat3 may lead to hepatotoxicity by mediating inflammation and cell proliferation. In conclusion, we proposed "quantity-weight-evidence" network toxicology in this study and used it to study the mechanism of DV-induced hepatotoxicity in rats. This study confirms the feasibility of this new methodology in toxicological evaluation and further improves the systematic evaluation of the safety of Chinese medicinal herbs.

Herbal Textual Research on Mori in Famous Classical Formulas / 中国实验方剂学杂志

Through consulting the ancient herbs， medical books and modern literature， this paper made textual research on the name， origin， producing area， quality evaluation， collection and processing of medicinal materials of Sang （Mori Folium， Mori Cortex， Mori Ramulus， Mori Fructus） in famous classical formulas， in order to provide a basis for the development of famous classical formulas containing medicinal materials of Sang. According to the research， Mori Folium and Mori Cortex were first used as medicines in Shengnong Bencaojing ， Mori Ramulus was first used as medicine in Jinxiaofang， and Mori Fructus was first used as medicine in Xinxiu Bencao. Before the Tang dynasty， there were Nyusang and Shansang. Since Tang dynasty， there were many sources of medicinal materials of Sang， including Baisang （Morus alba）， Jisang （M. australis）， Shansang （M. mongolica）， etc. According to textual research， the mainstream varieties were M. australis， M. alba and their cultivated varieties. In modern times， according to the relevant information and the Chinese Pharmacopoeia， M. alba is the original base. In ancient times， the origin of mulberry changed with the development of sericulture， mulberry has been widely planted since the Song dynasty. In the Ming and Qing dynasties， mulberry has been planted most in Jiangsu and Zhejiang. In modern times， they are mainly produced in Jiangsu， Zhejiang， Anhui， Hunan and other places. In recent years， due to the related policies and strategies such as "moving silkworms from east to west"， the center of silkworm breeding has gradually transferred to the west. As for the quality evaluation and harvesting and processing of mulberry medicinal materials， Most of the ancient and modern records of Mori Folium are the same. They are harvested after frost， and dried after removing impurities. The quality is better when the leaves are large and thick， yellowish green， holding prickly hands and undergoing frost. The harvesting period of Mori Cortex is slightly different in ancient and modern records. Ancient books record that it can be harvested all the year round， but in modern times， it is mostly harvested from late autumn to the next spring. The processing methods include removing soil and fibrous roots， scraping off yellow-brown rough skin， peeling off white skin and drying in the sun. The quality is better when they are white， thick， flexible， free of rough skin and full of powder. There are few records about the collection， processing and quality evaluation of Mori Ramulus and Mori Fructus in ancient Chinese herbal books. According to modern literature， Mori Ramulus is usually collected in late spring and early summer， with leaves removed， slightly dried， sliced while fresh， and dried in the sun. The best quality of Mori Ramulus is fine and tender with the yellow and white section. Mori Fructus is harvested from April to June when the fruit turns red， and dried in the sun， or slightly steamed and dried in the sun， and it is better to be big， dark purple， oily and thick. There are many processing methods of mulberry medicinal materials. Ancient books record stir frying， baking， burning and steaming of Mori Folium， in modern times， there is honey-roasted method， but most of them are used as raw products. In ancient materia medica， Mori Cortex has firing method， baking method， stir-frying method， honey-fried method， etc. In modern times， there are stir-fried and honey-fried methods， and most of them are used as raw products. Ancient books record that Mori Ramulus has cutting and frying methods， while modern ones have cutting， frying， wine-processed and bran-processed methods. Processing methods of Mori Fructus are consistent in ancient and modern times， and they are mostly dried after being cleaned or steamed. Based on the research results， it is suggested that M. alba should be selected as mulberry medicinal materials in the famous classical formulas， and appropriate medicinal parts and processing methods can be selected according to the indications of the famous classical formulas.

Herbal Textual Research on Menthae Haplocalycis Herba in Famous Classical Formulas / 中国实验方剂学杂志

In this paper， the name， origin， producing area and other aspects of Menthae Haplocalycis Herba in the famous classical formulas were carried out by consulting herbal literature， medical books， prescription books in the past dynasties and related modern documents. Through the textual research， it can be seen that the name of Bohe was used as the correct name in the mainstream of the past dynasties， and there were still multiple synonyms， most of which originated from the false transmission of dialectal accent， producing area and efficacy. There are many varieties recorded in the literature of the past dynasties such as Bohe， Longnao Bohe， Hubohe and Shibohe. According to the textual research， Bohe， Longnao Bohe and Yebohe are consistent with Mentha haplocalyx， whcih is the mainstream variety. Longnao Bohe is named for its form of producing area， Shibohe is Mosla chinensis， Daye Bohe is Agastache rugosa， and Nanbohe is M. crispata. Menthae Haplocalycis Herba has been widely planted since Tang dynasty. It was mainly grown in Jiangsu， Zhejiang， Jiangxi and Sichuan in Ming and Qing dynasties， and Jiangsu is the genuine production area. Its quality is best if it has dry body， no roots， many leaves， green color and strong smell. In ancient times， the stems and leaves of Menthae Haplocalycis Herba were often picked and dried in summer and autumn， which is basically the same as the records of modern times when the stems and leaves are luxuriant in summer and autumn， or when the flowers bloom to three rounds， they are picked in sunny days and cut in different times， and then dried in the sun or in the shade， and the raw products was often used as medicine in ancient and modern times. Before the Song dynasty， Menthae Haplocalycis Herba was recorded as pungent and warm. Until the Song dynasty， it was written as “extremely cool” in Lyuchanyan Bencao. It may have been thought in the early stage that it was similar to several warm herbs， such as Perilla frutescens， Stachys japonica， Elsholtzia ciliata and M. chinensis in appearance， all of which have the function of Xinsan， so it was recorded as warm. Since the Qing dynasty， Menthae Haplocalycis Herba has been recorded as cool property in the mainstream materia medica， Menthae Haplocalycis Herba recorded as pungent and cool in the 2020 edition of Chinese Pharmacopoeia， and its effect is to dissipate wind heat， clear the head， relieve the pharynx and so on， the records of efficacy in ancient and modern times are basically the same. Based on the research results， it is suggested that raw products of M. haplocalyx should be selected when developing the famous classical formulas containing Menthae Haplocalycis Herba.

Integrating network pharmacology and non-targeted metabolomics to explore the common mechanism of Coptis Categorized Formula improving T2DM zebrafish.

ETHNOPHARMACOLOGICAL RELEVANCE: Coptis Categorized Formula (CCF) is one of the core prescriptions in Treatise on Febrile Diseases. Its efficacy can be available not only in exogenous diseases but widely in various internal injuries and miscellaneous diseases. CCF (i.e., Huanglian Jiedu Decoction, Huanglian Ejiao Decoction, Dahuang Huanglian Xiexin Decoction, Gegen Qinlian Decoction) is different in composition, but they all play a favorable role in curative effect on type 2 diabetes mellitus (T2DM). Therefore, it is of great significance to reveal the common mechanism of CCF in treating T2DM. AIM OF THE STUDY: Based on network pharmacology and non-targeted metabolomics research strategy, the common mechanism of the CCF treating T2DM was discussed. MATERIALS AND METHODS: Firstly, Ultra-high performance liquid chromatography-quadrupole-time of flight/mass spectrometry was used to identify the chemical constituents of the CCF. Then, the targets of these chemical components were used for network pharmacology analysis associated with therapeutic effect. Finally, the diabetic zebrafish model was constructed to further verify the common mechanism of the CCF in treating T2DM. RESULTS: A total of 160 chemical compositions were identified and 16 of them were common chemical compositions of the four CCF, including berberine, baicalin, coptisine and so forth. Network pharmacology results showed that Dipeptidyl peptidase (DPP)-4, cysteinyl aspartate specific proteinase (CASP)3, nitric oxide synthase (NOS)2, NOS3, and other 37 targets were common targets of CCF, and advanced glycation end products (AGE)-receptor of advanced glycation end products (RAGE) signaling pathway in diabetic complications, mitogen-activated protein kinase (MAPK) signaling pathway and hypoxia inducible factor (HIF)-1 signaling pathway were critical pathways of four CCF in the treatment of T2DM. CCF can lessen the blood glucose of diabetic zebrafish. The contents of 25 differential metabolites in diabetic zebrafish were altered. These metabolites were mainly related to phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, arachidonic acid metabolism, sphingolipid metabolism, and tyrosine metabolism. CONCLUSION: Our research shows that the common mechanism of CCF in improving T2DM is as follows: berberine, baicalin, coptisine and other chemical components can directionally regulate DPP-4, CASP3, NOS2, NOS3 and other targets, which are mediated by AGE-RAGE signaling pathway in diabetic complications, MAPK signaling pathway and HIF-1 signaling pathway. The content of endogenous metabolites such as L-valine and L-sorbitose changes, and further regulates the metabolism of amino acid metabolism, lipid metabolism, purine metabolism, sphingosine metabolism and arachidonic acid metabolism, so as to play a significant role in regulating glycolipid metabolism, improving insulin resistance, inhibiting cell apoptosis, anti-oxidation and anti-inflammation, and finally ameliorating T2DM.

Xi-Xian-Tong-Shuan capsule alleviates vascular cognitive impairment in chronic cerebral hypoperfusion rats by promoting white matter repair, reducing neuronal loss, and inhibiting the expression of pro-inflammatory factors.

BACKGROUND: While the number of cases of vascular cognitive impairment caused by chronic cerebral hypoperfusion (CCH) has been increasing every year, there are currently no clinically effective treatment methods. At present, Xi-Xian-Tong-Shuan capsule is predominantly used in patients with acute cerebral ischemia; however, its protective effect on CCH has rarely been reported. OBJECTIVE: To explore the underlying mechanisms by which Xi-Xian-Tong-Shuan capsule alleviates cognitive impairment caused by CCH. METHODS: A model of CCH was established in specific-pathogen-free (SPF)-grade male Sprague-Dawley (SD) rats using bilateral common carotid artery occlusion (BCCAO). Xi-Xian-Tong-Shuan capsules were intragastrically administered for 42 days after the BCCAO surgery. We then assessed for changes in cognitive function, expression levels of pro-inflammatory factors, and coagulation function as well as for the presence of white matter lesions and neuronal loss. One-way ANOVA and Tukey's test were used to analyze the experimental data. RESULTS: The rats showed significant cognitive dysfunction after the BCCAO surgery along with white matter lesions, a loss of neurons, and elevated levels of inflammatory factors, all of which were significantly reversed after intervention with Xi-Xian-Tong-Shuan capsules. CONCLUSION: Xi-Xian-Tong-Shuan capsules can ameliorate vascular cognitive impairment in CCH rats by preventing damage of white matter, reducing neuronal loss, and inhibiting the expression of pro-inflammatory factors. Our study provides a new reference for the clinical treatment of chronic cerebral ischemia with Xi-Xian-Tong-Shuan capsules.

The Research on the Treatment of Metastatic Skin Cutaneous Melanoma by Huanglian Jiedu Decoction Based on the Analysis of Immune Infiltration Analysis.

OBJECTIVE: To explore the potential mechanism of Huanglian Jiedu Decoction (HJD) treatment and prevention of metastatic Cutaneous Melanoma (CM) occurrence and metastasis based on network pharmacological methods and immune infiltration analysis. METHODS: The GEO database was used to obtain metastatic CM disease targets, the TCMSP database and the HERB database were used to obtain HJD action targets, core genes were screened by protein interaction network, and the potential mechanism of HJD in the treatment of metastatic CM was explored by enrichment analysis, prognostic analysis and immune infiltration analysis. RESULTS: HJD treatment of metastatic CM involved 60 targets, enrichment analysis showed that HJD treatment of metastatic CM involved Chemokine signaling pathway, NF-kappa B signaling pathway, and Fluid shear stress and atherosclerosis, etc. Prognostic analysis revealed that HJD had a certain ability to improve the prognosis of metastatic CM patients. Immune infiltration analysis showed that HJD could inhibit the immune cell infiltration of metastatic CM patients by acting on related targets. CONCLUSIONS: Our study identified the potential mechanism of HJD in the treatment of metastatic CM through network pharmacology, and revealed the mechanism of HJD in the prevention of Skin Cutaneous Melanoma metastasis through immune infiltration analysis and prognostic analysis.

Diversity and function of rhizosphere microorganisms between wild and cultivated medicinal plant Glycyrrhiza uralensis Fisch under different soil conditions.

Glycyrrhiza uralensis Fisch is a widely cultivated traditional Chinese medicine plant. In the present study, culture-independent microbial diversity analysis and functional prediction of rhizosphere microbes associated with wild and cultivated G. uralensis Fisch plant (collected from two locations) were carried. Soil physicochemical parameters were tested to assess their impact on microbial communities. A total of 4428 OTUs belonging to 41 bacterial phyla were identified. In general, cultivated sample sites were dominated by Actinobacteria whereas wild sample sites were dominated by Proteobacteria. The alpha diversity analysis showed the observed species number was higher in cultivated soil samples when compared with wild soil samples. In beta diversity analysis, it was noticed that the weighted-unifrac distance of two cultivated samples was closer although the samples were collected from different regions. Functional annotation based on PICRUST and FAPROTAX showed that the nitrogen metabolism pathway such as nitrate reduction, nitrogen fixation, nitrite ammonification, and nitrite respiration were more abundant in rhizosphere microorganisms of wild G. uralensis Fisch. These results also correlate in redundancy analysis results which show correlation between NO3--N and wild samples, which indicated that nitrogen nutrition conditions might be related to the quality of G. uralensis Fisch.

[Herbal textual research on Rubi Fructus].

Rubi Fructus is a commonly used traditional Chinese medicine. The origin of Rubi Fructus is the dried fruit of Rubus chingii, a plant of the family Rosaceae, according to the 2015 edition of Chinese pharmacopoeia. There are some differences in the plant origin of Rubi Fructus in ancient herbal literature, to trace back its sources, we conducted a textual research on its origin, producing areas, quality evaluation, processing and concocting, properties, tastes and efficacy etc. based on the records of ancient herbal literatures and combined with plant morphology and related investigation. RESULTS:: showed that the variety of Rubi Fructus was more complex among ancient herbal literature, including R. coreanus, R. hirsutus, R. corchorifolius, R. foliolosus and other mixed varieties. Most scholars believe that the R. chingii has not been recorded in ancient herbal literature, while R. chingii was recorded as early as the Ming Dynasty in Compendium of materia medica through our textual research. Ancient Chinese herbs recorded that Rubi Fructus was mostly produced in Hubei, Shandong, Shanxi and Jiangsu provinces, while R. chingii mainly produced in Anhui, Jiangsu, Zhejiang, Jiangxi, Fujian and other provinces nowadays. Also, it was recorded that Rubi Fructus harvested in wheat field during May were the best. Besides, R. chingii with big, full, grain integrate, firm, yellow and green color, sour taste and impurity free possess the best quality in the contemporary. The ancient records of processing and concocting, properties, tastes and efficacy were basically the same as modern ones.These results provide the basis for the correct utilization and further development of Rubi Fructus.

Textual Research on Visci Herba / 中国实验方剂学杂志

Visci Herba， a commonly used Chinese medicinal， was often mistaken as Taxilli Herba in ancient Chinese materia medica. The two Chinese medicinals have often been confused even in present clinical practice， and their origins are difficult to be identified. Hence， it is necessary to carry out systematic and in-depth textual research. This paper explored the origin， producing area， quality evaluation， harvesting and processing， property， flavor， and efficacy of Visci Herba based on ancient Chinese materia medica of the past dynasties and modern plant morphology， so as to provide evidence for the development and utilization of Visci Herba. The findings demonstrated that Visci Herba was mostly recorded as Taxilli Herba until the name of Visci Herba appeared in the Tang Dynasty. The records of the two Chinese medicinals could be traced back to the Song Dynasty. Visci Herba and Taxilli Herba were officially listed as two different Chinese medicinals in the 1977 edition Chinese Pharmacopoeia for the first time， where the origin of Visci Herba was determined to be Viscum coloratum（Komar.）Nakai. According to the ancient Chinese materia medica， V. coloratum was mainly distributed in Lingbao， Henan province and Xuzhou， Jiangsu province and now it mostly grows in Northeast China and North China. In ancient times， Visci Herba with deep yellow cross-section and sticky fruit juice on the tree was preferred， which was often harvested on 3， March in spring， dried in the shade， grinded together with the roots， branches， stems， and leaves using the copper file， and preserved away from fire. By contrast， it is now usually harvested from winter to the next spring and then cut into sections for drying after the removal of thick stems， or dried after being steamed. As described in ancient Chinese materia medica， Visci Herba， bitter and sweet in flavor， neutral in property， possessed the effects of dispelling rheumatism， tonifying kidney， strengthening sinews and bones， benefiting blood vessels， and preventing abortion， basically consistent with its modern functions of dispelling rheumatism， tonifying liver and kidney， strengthening sinews and bones， and preventing abortion.