1.Exploring Mechanism of Hei Xiaoyaosan Regulating PI3K/Akt Pathway to Improve Learning and Memory Ability of Insomnia Rats with Liver Depression Syndrome Based on Transcriptomics
Jiamin LIU ; Yale WANG ; Hai HUANG ; Yue LI ; Xin FAN ; Pengpeng LIANG ; Shizhao ZHANG ; Mei YAN ; Guiyun LI ; Hongyan WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):114-125
ObjectiveBased on transcriptomics, to explore the mechanism of Hei Xiaoyaosan regulating the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway to improve the learning and memory ability of insomnia rats with liver depression syndrome. MethodsSixty 8-week-old male SD rats were randomly divided into the blank group, model group, eszopiclone group (0.09 mg·kg-1), and low, medium, and high dose groups of Hei Xiaoyaosan (3.82, 7.65, 15.30 g·kg-1), with ten rats in each group. Except for the blank group, the other groups were induced insomnia rat model with liver depression by chronic restraint, tail clamping stimulation and intraperitoneal injection of p-chlorophenylalanine (PCPA). Each treatment group received intragastric administration according to the specified dosage, once a day for 14 consecutive days. The pentobarbital sodium cooperative sleep test, open field test, and Morris water maze test were used to test the sleep quality, depressive-like behavior, and learning and memory abilities of rats. Additionally, enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nitric oxide (NO) in hippocampus. Hematoxylin-eosin (HE) staining was performed to observe pathological changes of the hippocampal tissue, while terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) was used to evaluate apoptosis of hippocampal neurons. Transcriptomic sequencing technology was employed to identify differentially expressed genes in hippocampus between the model group and the blank group, as well as between the medium-dose group of Hei Xiaoyaosan and the model group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on the intersecting genes. Subsequently, the enriched key genes and signaling pathways were analyzed and verified. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was utilized to assess the mRNA expression levels of phosphatase and tensin homolog (PTEN), B-cell lymphoma-2 (Bcl-2)-like protein 11 (BCL2L11), and mitogen-activated protein kinase 1 (MAPK1) in hippocampus, and Western blot was employed to evaluate the protein expressions of PI3K, phosphorylation (p)-PI3K, Akt, p-Akt, Bcl-2, Bcl-2-associated X protein (Bax), and cleaved Caspase-3 in the same tissue. ResultsCompared with the blank group, the model group exhibited a reduction in body weight, an increase in sleep latency, and a decrease in sleep duration (P<0.01). Additionally, rats showed obvious depression-like behavior, and their learning and memory abilities decreased. Furthermore, the contents of 5-HT, GABA, NO, BDNF and GDNF in hippocampus decreased (P<0.01). Histological examination revealed a disorganized cell arrangement in the CA1 region of the hippocampus, characterized by irregular cell shapes, a reduced cell count, deeply stained and pyknotic nuclei, increased vacuolar degeneration, and an elevated apoptosis rate (P<0.01). Compared with the model group, the body weight of the high and medium dose groups of Hei Xiaoyaosan increased, the sleep latency shortened and the sleep time prolonged (P<0.05, P<0.01). Additionally, depression-like behavior and learning and memory abilities of rats were significantly improved, the levels of 5-HT, GABA, NO, BDNF and GDNF in the hippocampus increased (P<0.05, P<0.01). These interventions also ameliorated pathological damage in the hippocampal CA1 area and reduced the apoptosis of hippocampal neurons (P<0.01). Transcriptomic sequencing results indicated that Hei Xiaoyaosan might exert a therapeutic effect by regulating PI3K/Akt pathway through key mRNAs such as PTEN, BCL2L11, and MAPK1. The roles of these key mRNAs and proteins within PI3K/Akt pathway were further validated. In comparison to the blank group, the expression levels of PTEN, BCL2L11 and MAPK1 mRNA in the hippocampus of rats in the model group were increased (P<0.01), while the protein expression levels of p-PI3K, p-Akt and Bcl-2 were decreased (P<0.01), and the protein expression levels of PTEN, Bax and cleaved Caspase-3 were increased (P<0.01). Compared with the model group, the high-dose and medium-dose groups of Hei Xiaoyaosan could down-regulate the expressions of PTEN, BCL2L11 and MAPK1 mRNAs (P<0.01), up-regulate the expressions of p-PI3K, p-Akt and Bcl-2 proteins (P<0.01), and down-regulate the protein expressions of PTEN, Bax and cleaved Caspase-3 (P<0.05, P<0.01). ConclusionHei Xiaoyaosan may regulate PI3K/Akt signaling pathway by down-regulating expressions of key genes such as PTEN, BCL2L11 and MAPK1, and thus improve the learning and memory abilities of insomnia rats with liver depression syndrome.
2.Comparative study of five physical disability-related standards in China based on ICF and ICD-11
Haiyan YE ; Yaru YANG ; Zhuoying QIU ; Zhongyan WANG ; Di CHEN ; Guiyun SONG ; Fangyong WANG ; Yifan TIAN ; Ye LIU
Chinese Journal of Rehabilitation Theory and Practice 2025;31(7):772-780
Objective To systematically compare five physical disability-related standards widely used in the fields of insurance,work related injury,forensic appraisal,and disability evaluation.The comparison covers definition,coding,classi-fication,grading,and assessment methods to reveal each standard's framework and methodology,and to provide evidence for integrating and optimizing physical disability standards.Methods Based on International Classification of Functioning,Disability and Health(ICF)and International Classifica-tion of Diseases,11th Revision(ICD-11),we analyzed and compared five physical disability-related standards,including Disability Assessment and Code for Life Insurance(Insurance Standard),Grading of Disability due to Human Body Injury(Judicial Standard),Standard for Identify Work Ability—Gradation of Disability Caused by Work-related Injuries and Occupational Diseases(Work Injury Standard),Standard for Assessment of Disability Grades of Military Personnel(Military Standard),and Classification and Grading Criteria of Disability(Disabili-ty Classification Standard).We examined definition,coding systems,classification,grading scales and assess-ment.Results According to the ICF framework,at the level of body functions and structures,physical disability mainly in-volved neuromusculoskeletal and movement related functions(b7),including joint and bone functions(b710-b729),muscle functions(b730-b749),and movement functions(b750-b789).At the activities and participation level,it primarily concerned mobility(d410-d499)and self care(d510-d599).Related conditions were commonly found in Certain infectious or parasitic diseases(01),Diseases of the nervous system(08),Diseases of the muscu-loskeletal system or connective tissue(15),Developmental anomalies(20),and Injury,poisoning or certain other consequences of external causes(22),etc.Among the five standards,only Disability Classification Standard ex-plicitly defined physical disability,yet its terminology did not followed ICF categories.Regarding coding,only the Insurance Standard adopted ICF codes;and the other four used self devised coding systems.The sub catego-ries in these physical disability standards did not fully cover all ICF categories relevant to physical disability.In terms of grading,Disability Classification Standard used a four level grading,whereas the other four standards employed ten level grading.Concerning assessment,all five emphasized evaluation of body structures and func-tions and involved daily activity capacity;however,WHO Disability Assessment Schedule(WHODAS 2.0)for comprehensive functional assessment,had not been proposed for evaluation of general functioning.Conclusion China has developed different physical disability standards to meet diverse needs.There are differences in definition,structure and content.It is necessary to revise and refine the relevant components of physical disabili-ty,including definition,classification,grading,coding and assessment,based on ICF and ICD-11,and to intro-duce WHODAS 2.0 as a comprehensive functional assessment tool.
3.Huanglian Jiedu decoction combined with Xijiao Dihuang decoction for the treatment of psoriasis via influencing fibroblast activation-mediated keratinocyte proliferation: a mechanistic study
Youhua PENG ; Guiyun GAO ; Chao LIU ; Jinglin LI ; Mengyao ZHANG ; Jing DAI ; Yao CHEN ; Junqi LIU ; Xudong WANG
Chinese Journal of Dermatology 2025;58(11):1064-1074
Objective:To explore the mechanisms of action of Huanglian Jiedu decoction combined with Xijiao Dihuang decoction (HLJDT-XJDH) in regulating fibroblasts in the treatment of psoriasis. Methods:A mouse model of psoriasis was established by topical application of imiquimod 5% cream on the shaved back; HLJDT-XJDH at different doses of 7.7 and 30.6 g/kg was administered via gavage for intervention, and methotrexate (2 mg/kg) served as a positive control; after 7 days, the severity of skin lesions was assessed using the psoriasis area and severity index (PASI), while histopathological changes of skin tissues were evaluated using hematoxylin-eosin (HE) staining and Baker scoring. For in vitro experiments, fibroblasts were divided into a control group, a model group, a low-dose (5% drug-containing serum) intervention group, and a high-dose (20% drug-containing serum) intervention group; cells in the control group were cultured with 20% normal rat serum for 24 hours; in the model group, cells cultured with 20% normal rat serum were stimulated with 5 ng/ml tumor necrosis factor (TNF) -α and 50 ng/ml interleukin (IL) -17A for 24 hours to mimic fibroblasts during the occurrence of psoriasis; cells in the low- and high-dose intervention groups received the same stimulation as the model group, and were cultured for 24 hours with 5% and 20% HLJDT-XJDH-containing serum, respectively, but not with the 20% normal rat serum. After the above treatment, these cells were co-cultured with keratinocytes (HaCaT cells) using a Transwell system. In addition, on the basis of the control group, fibroblasts were divided into the model group, 20% drug-containing serum intervention group, and 20% drug-containing serum intervention + OE-SFRP2 group; TNF-α and IL-17A were used to stimulate the cells to simulate the psoriatic state; the treatment in the 20% drug-containing serum intervention group was carried out as previously described; in the 20% drug-containing serum intervention + OE-SFRP2 group, cells were transfected with the vector for 48 hours to establish an overexpression model, followed by culture with 20% drug-containing serum for 24 hours, without co-culture with HaCaT cells.. Cell counting kit-8 (CCK-8) assay was performed to assess cell viability, flow cytometry to measure apoptosis rates, enzyme-linked immunosorbent assay (ELISA) to detect levels of inflammatory cytokines (TNF-α, IL-1β, IL-6) as well as chemokine ligand (CXCL) 1 and CXCL12 in mouse serum or cell culture supernatant, qPCR to determine the mRNA expression of inflammatory cytokines, chemokines, cell cycle- and proliferation-related factors, as well as SFRP2 in mouse skin tissues or cells, and Western blot analysis to determine the protein expression of SFRP2, Wnt3a, and β-catenin in fibroblasts. One-way analysis of variance was employed for intergroup comparisons, and post-hoc analysis was conducted using Tukey's test. Results:In vivo mouse experiments showed that compared with the normal control group, the model group exhibited typical psoriatic characteristics in skin morphology, including significant inflammatory infiltration in skin tissues and marked epidermal thickening; compared with the normal control group, the serum levels of TNF-α (531.16 ± 28.27 pg/ml vs. 239.58 ± 10.39 pg/ml), IL-1β (111.40 ± 5.16 pg/ml vs. 80.35 ± 3.87 pg/ml), and IL-6 (109.17 ± 4.84 pg/ml vs. 71.73 ± 2.04 pg/ml) significantly increased in the model group, along with their mRNA expression levels in mouse skin tissues (all P < 0.001) ; compared with the model group, the treatment group showed alleviated psoriatic manifestations, and significant reductions in the levels of inflammatory factors TNF-α (low-dose, high-dose, and positive control groups: 420.80 ± 29.30 pg/ml, 322.33 ± 9.40 pg/ml, 322.97 ± 12.16 pg/ml, respectively), IL-1β (98.69 ± 4.49 pg/ml, 89.02 ± 1.56 pg/ml, 88.88 ± 2.08 pg/ml, respectively), and IL-6 (94.07 ± 3.76 pg/ml, 80.54 ± 3.30 pg/ml, 83.21 ± 3.18 pg/ml, respectively), as well as in their mRNA expression levels (all P < 0.001). In in vitro fibroblast experiments, compared with the control group, the model group exhibited a significant elevation in the supernatant levels of IL-1β (126.42 ± 3.56 pg/ml vs. 34.81 ± 0.44 pg/ml), IL-6 (459.44 ± 9.35 pg/ml vs. 115.51 ± 7.26 pg/ml), CXCL1 (2 434.88 ± 127.63 pg/ml vs. 762.85 ± 30.60 pg/ml) and CXCL12 (3 542.14 ± 35.86 pg/ml vs. 2 095.86 ± 45.12 pg/ml), the expression levels of their mRNAs (all P < 0.001), as well as the protein expression levels of SFRP2, Wnt3a, and β-catenin; after intervention with HLJDT-XJDH-containing serum, all the above indices significantly decreased (all P < 0.001). However, when 20% drug-containing serum intervention was administered simultaneously, the expression of inflammatory factors and chemokines in fibroblasts was significantly higher in the SFRP2 overexpression group than in the non-overexpression group (all P < 0.01). When fibroblasts were co-cultured with HaCaT cells, the model group showed significantly increased cell viability but a decreased apoptosis rate of HaCaT cells compared with the control group, while the low- and high-dose intervention groups showed significantly decreased cell viability but increased apoptosis rates of HaCaT cells compared with the model group (all P < 0.05) . Conclusion:HLJDT-XJDH may exert therapeutic effects in psoriasis by downregulating the SFRP2/Wnt/β-catenin signaling pathway, thereby inhibiting fibroblast activation and inflammatory process, which subsequently suppresses the proliferation of keratinocytes and the activation of inflammatory cells.
4.Experience of Using Charcoal-Processed Traditional Chinese Medicine in the Treatment of Gynecological Hemorrhagic Disorders
Xiaolan WU ; Zhaoling YOU ; Guiyun WANG ; Kailing WANG ; Xiaojuan YE ; Lingyu LIAO ; Yueheng LI ; Huiping LIU
Journal of Traditional Chinese Medicine 2025;66(3):308-311
Charcoal-processed traditional Chinese herbal medicine has various therapeutic effects, including astringing, hemostasis, anti-diarrhea, clearing heat, tonifying, and warming the interior. This paper summarizes the clinical application features, compatible experiences, dosages, and precautions for over 20 types of charcoal-processed herbal medicine in the treatment of gynecological bleeding disorders caused by dysfunctions such as dysfunctional uterine bleeding, endometriosis, uterine incision pseudocavity, and vaginal bleeding resulting from threatened miscarriage. The charcoal-processed herbal medicine include Huangqin (Scutellaria Baicalensis) Charcoal, Dahuang (Rheum Palmatum) Charcoal, Cebai (Platycladus Orientalis) Charcoal, Diyu (Sanguisorba Officinalis) Charcoal, Daji (Cirsium Setosum) Charcoal, Xiaoji (Cirsium Japonicum) Charcoal, Shengdi (Rehmannia Glutinosa) Charcoal, Aiye (Artemisia Argyi) Charcoal, Paojiang (Zingiber Officinale) Charcoal, Xuduan (Dipsacus Asper) Charcoal, Duzhong (Eucommia Ulmoides) Charcoal, Qiancao (Rubia Cordifolia) Charcoal, Puhuang (Typha Angustifolia) Charcoal, Shanzha (Crataegus Pinnatifida) Charcoal, Jingjie (Schizonepeta Tenuifolia) Charcoal, Xueyu (Carthamus Tinctorius) Charcoal, Zonglyu (Areca Catechu) Charcoal, Wumei (Prunus Mume) Charcoal, Shudahuang (Rheum Officinale) Charcoal, Lianfang (Nymphaea Alba) Charcoal, Mianmaguanzhong (Clematis Armandii) Charcoal, and Oujie (Nelumbo Nucifera) Charcoal.
5.Effect of Bushen Huoxue Prescription in Regulating PINK1/Parkin Pathway in Rat Model of Premature Ovarian Failure
Kailing WANG ; Yichen JING ; Guiyun WANG ; Yueheng LI ; Huiping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):150-158
ObjectiveTo investigate the mechanism of action of Bushen Huoxue prescription (BSHXP) in regulating premature ovarian failure in rats through the PTEN-induced kinase 1 (PINK1)/Parkinson's protein (Parkin) signaling pathway-mediated mitophagy. MethodsA total of 48 rats were randomly divided into a blank group consisting of eight rats, while the remaining 40 rats underwent modeling. The modeling group was intraperitoneally injected with 4 mg·kg-1 cisplatin solution, followed by a second injection one week later, for a total of two injections. The estrous cycle was observed through vaginal smears for 14 consecutive days to determine whether the modeling was successful. The successfully modeled rats were randomly divided into a model group, groups receiving low, medium, and high doses of BSHXP at 9.72, 19.44, and 38.88 g·kg-1·d-1 (BSHXP-L, BSHXP-M, and BSHXP-H groups), and a positive control group treated with estradiol valerate (0.09 mg·kg-1·d-1), for 21 consecutive days. The body weight of the rats was measured weekly. After the final administration, rats were anesthetized, and their blood and ovaries were collected. The ovarian weight was measured. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2). Assay kits were used to measure the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the rat serum. Hematoxylin-eosin (HE) staining was used to observe the morphological changes in the ovaries. Immunohistochemistry (IHC) was performed to detect microtubule autophagy-related protein 1 light chain 3B(LC3B) protein expression in ovarian tissue, and electron microscopy was employed to examine the mitochondrial and autophagosome changes in the rat ovaries. Western blot was used to detect the protein expression of PINK1, Parkin, LC3B, and p62. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of PINK1, Parkin, LC3B, and p62 in ovarian tissue. ResultsCompared with the blank group, the model group showed significant reductions in body weight, weight gain, and ovarian weight (P<0.01), along with decreased serum AMH and E2 levels (P<0.01), while FSH and LH levels were increased (P<0.01). Serum MDA levels were significantly increased (P<0.01), and SOD levels were significantly reduced (P<0.01). The ovarian tissue structure was disordered, and the zona pellucida was wrinkled into an irregular acidophilic annular object, accompanied by an increased number of closed follicles. Electron microscopy showed mitochondrial swelling, unclear structure, and no obvious autophagosomes and autolysosome structures. The proteins and mRNA expression levels of PINK1, Parkin, LC3B, and p62 in the ovarian tissue were significantly reduced (P<0.01). Compared with the model group, all treatment groups showed varying degrees of increases in body weight and ovarian weight (P<0.05, P<0.01). Except for the BSHXP-L group, all treatment groups showed increased body weight gain (P<0.01). All treatment groups showed significantly increased serum AMH and decreased FSH levels (P<0.01). Except for the BSHXP group, all treatment groups showed varying degrees of increase and decrease in serum E2 and LH levels (P<0.05, P<0.01). All treatment groups showed reduced serum MDA levels (P<0.01), while the BSHXP-M, BSHXP-H, and the positive control groups demonstrated improved serum SOD levels (P<0.05, P<0.01). All treatment groups showed an increased number of follicles at all stages, visible mature follicles, and a decreased number of closed follicles. Electron microscopy showed relieved mitochondrial swelling, morphology close to normal, clear structure, and visible formation of autolysosomes in all treatment groups. Additionally, the protein and mRNA expression levels of PINK1, Parkin, LC3B, and p62 in ovarian tissue were significantly increased (P<0.05, P<0.01). ConclusionBSHXP may improve ovarian function in rats with premature ovarian failure by regulating the PINK1/Parkin signaling pathway, activating mitochondrial autophagy, and reducing oxidative damage.
6.Effect of Bushen Huoxue Prescription in Regulating PINK1/Parkin Pathway in Rat Model of Premature Ovarian Failure
Kailing WANG ; Yichen JING ; Guiyun WANG ; Yueheng LI ; Huiping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):150-158
ObjectiveTo investigate the mechanism of action of Bushen Huoxue prescription (BSHXP) in regulating premature ovarian failure in rats through the PTEN-induced kinase 1 (PINK1)/Parkinson's protein (Parkin) signaling pathway-mediated mitophagy. MethodsA total of 48 rats were randomly divided into a blank group consisting of eight rats, while the remaining 40 rats underwent modeling. The modeling group was intraperitoneally injected with 4 mg·kg-1 cisplatin solution, followed by a second injection one week later, for a total of two injections. The estrous cycle was observed through vaginal smears for 14 consecutive days to determine whether the modeling was successful. The successfully modeled rats were randomly divided into a model group, groups receiving low, medium, and high doses of BSHXP at 9.72, 19.44, and 38.88 g·kg-1·d-1 (BSHXP-L, BSHXP-M, and BSHXP-H groups), and a positive control group treated with estradiol valerate (0.09 mg·kg-1·d-1), for 21 consecutive days. The body weight of the rats was measured weekly. After the final administration, rats were anesthetized, and their blood and ovaries were collected. The ovarian weight was measured. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2). Assay kits were used to measure the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the rat serum. Hematoxylin-eosin (HE) staining was used to observe the morphological changes in the ovaries. Immunohistochemistry (IHC) was performed to detect microtubule autophagy-related protein 1 light chain 3B(LC3B) protein expression in ovarian tissue, and electron microscopy was employed to examine the mitochondrial and autophagosome changes in the rat ovaries. Western blot was used to detect the protein expression of PINK1, Parkin, LC3B, and p62. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of PINK1, Parkin, LC3B, and p62 in ovarian tissue. ResultsCompared with the blank group, the model group showed significant reductions in body weight, weight gain, and ovarian weight (P<0.01), along with decreased serum AMH and E2 levels (P<0.01), while FSH and LH levels were increased (P<0.01). Serum MDA levels were significantly increased (P<0.01), and SOD levels were significantly reduced (P<0.01). The ovarian tissue structure was disordered, and the zona pellucida was wrinkled into an irregular acidophilic annular object, accompanied by an increased number of closed follicles. Electron microscopy showed mitochondrial swelling, unclear structure, and no obvious autophagosomes and autolysosome structures. The proteins and mRNA expression levels of PINK1, Parkin, LC3B, and p62 in the ovarian tissue were significantly reduced (P<0.01). Compared with the model group, all treatment groups showed varying degrees of increases in body weight and ovarian weight (P<0.05, P<0.01). Except for the BSHXP-L group, all treatment groups showed increased body weight gain (P<0.01). All treatment groups showed significantly increased serum AMH and decreased FSH levels (P<0.01). Except for the BSHXP group, all treatment groups showed varying degrees of increase and decrease in serum E2 and LH levels (P<0.05, P<0.01). All treatment groups showed reduced serum MDA levels (P<0.01), while the BSHXP-M, BSHXP-H, and the positive control groups demonstrated improved serum SOD levels (P<0.05, P<0.01). All treatment groups showed an increased number of follicles at all stages, visible mature follicles, and a decreased number of closed follicles. Electron microscopy showed relieved mitochondrial swelling, morphology close to normal, clear structure, and visible formation of autolysosomes in all treatment groups. Additionally, the protein and mRNA expression levels of PINK1, Parkin, LC3B, and p62 in ovarian tissue were significantly increased (P<0.05, P<0.01). ConclusionBSHXP may improve ovarian function in rats with premature ovarian failure by regulating the PINK1/Parkin signaling pathway, activating mitochondrial autophagy, and reducing oxidative damage.
7.Study on the antagonistic activity of Lactococcus garvieae SHAMU-LG6 against Staphylococcus
Shengnan Weng ; Guiyun Leng ; Ying Liu ; Yawu Wang ; Xin Li ; Jie Yao ; Qiang Zhou ; Wei Tang
Acta Universitatis Medicinalis Anhui 2025;60(2):195-200
Objective :
To investigate the antagonistic activity of Lactococcus garvieae SHAMU-LG6 against Staphy- lococcus .
Methods :
VITEK 2 GP identification card , Microflex LT MALDI-TOF mass spectrometer and 16S rDNA amplification sequencing were used to identify the strain species . The antagonistic activity of L. garvieae SHAMU- LG6 against different Staphylococcus was detected by Oxford cup method for bacterial inhibition ; the antimicrobial active components were preliminarily isolated and purified by adsorption on XAD16 nonionic macroporous resin , gradient ethanol elution and rotary evaporation drying.
Results :
L. garvieae SHAMU-LG6 exhibited potent antago- nistic effect against methicillin-resistant Staphylococcus aureus , methicillin-susceptible S. aureus , S. epidermidis , S. saprophyticus , S. lugdunensis , S. hominis , S. capitis and S. warneri , with inhibitory indices of 3 . 3 , 3 . 0 , 4. 3 , 2. 0 , 4. 0 , 3 . 5 , 3 . 8 , and 3 . 5 , respectively. The antimicrobial active components produced by L. garvieae SHAMU-LG6 were mainly present in 70% and 80% ethanol eluates .
Conclusion
L. garvieae SHAMU-LG6 ex- hibits a potent antagonistic effect on Staphylococcus , and the antimicrobial active components produced by it are ex- pected to be a lead compound for the development of novel antimicrobial agents .
8.Huanglian Jiedu decoction combined with Xijiao Dihuang decoction for the treatment of psoriasis via influencing fibroblast activation-mediated keratinocyte proliferation: a mechanistic study
Youhua PENG ; Guiyun GAO ; Chao LIU ; Jinglin LI ; Mengyao ZHANG ; Jing DAI ; Yao CHEN ; Junqi LIU ; Xudong WANG
Chinese Journal of Dermatology 2025;58(11):1064-1074
Objective:To explore the mechanisms of action of Huanglian Jiedu decoction combined with Xijiao Dihuang decoction (HLJDT-XJDH) in regulating fibroblasts in the treatment of psoriasis. Methods:A mouse model of psoriasis was established by topical application of imiquimod 5% cream on the shaved back; HLJDT-XJDH at different doses of 7.7 and 30.6 g/kg was administered via gavage for intervention, and methotrexate (2 mg/kg) served as a positive control; after 7 days, the severity of skin lesions was assessed using the psoriasis area and severity index (PASI), while histopathological changes of skin tissues were evaluated using hematoxylin-eosin (HE) staining and Baker scoring. For in vitro experiments, fibroblasts were divided into a control group, a model group, a low-dose (5% drug-containing serum) intervention group, and a high-dose (20% drug-containing serum) intervention group; cells in the control group were cultured with 20% normal rat serum for 24 hours; in the model group, cells cultured with 20% normal rat serum were stimulated with 5 ng/ml tumor necrosis factor (TNF) -α and 50 ng/ml interleukin (IL) -17A for 24 hours to mimic fibroblasts during the occurrence of psoriasis; cells in the low- and high-dose intervention groups received the same stimulation as the model group, and were cultured for 24 hours with 5% and 20% HLJDT-XJDH-containing serum, respectively, but not with the 20% normal rat serum. After the above treatment, these cells were co-cultured with keratinocytes (HaCaT cells) using a Transwell system. In addition, on the basis of the control group, fibroblasts were divided into the model group, 20% drug-containing serum intervention group, and 20% drug-containing serum intervention + OE-SFRP2 group; TNF-α and IL-17A were used to stimulate the cells to simulate the psoriatic state; the treatment in the 20% drug-containing serum intervention group was carried out as previously described; in the 20% drug-containing serum intervention + OE-SFRP2 group, cells were transfected with the vector for 48 hours to establish an overexpression model, followed by culture with 20% drug-containing serum for 24 hours, without co-culture with HaCaT cells.. Cell counting kit-8 (CCK-8) assay was performed to assess cell viability, flow cytometry to measure apoptosis rates, enzyme-linked immunosorbent assay (ELISA) to detect levels of inflammatory cytokines (TNF-α, IL-1β, IL-6) as well as chemokine ligand (CXCL) 1 and CXCL12 in mouse serum or cell culture supernatant, qPCR to determine the mRNA expression of inflammatory cytokines, chemokines, cell cycle- and proliferation-related factors, as well as SFRP2 in mouse skin tissues or cells, and Western blot analysis to determine the protein expression of SFRP2, Wnt3a, and β-catenin in fibroblasts. One-way analysis of variance was employed for intergroup comparisons, and post-hoc analysis was conducted using Tukey's test. Results:In vivo mouse experiments showed that compared with the normal control group, the model group exhibited typical psoriatic characteristics in skin morphology, including significant inflammatory infiltration in skin tissues and marked epidermal thickening; compared with the normal control group, the serum levels of TNF-α (531.16 ± 28.27 pg/ml vs. 239.58 ± 10.39 pg/ml), IL-1β (111.40 ± 5.16 pg/ml vs. 80.35 ± 3.87 pg/ml), and IL-6 (109.17 ± 4.84 pg/ml vs. 71.73 ± 2.04 pg/ml) significantly increased in the model group, along with their mRNA expression levels in mouse skin tissues (all P < 0.001) ; compared with the model group, the treatment group showed alleviated psoriatic manifestations, and significant reductions in the levels of inflammatory factors TNF-α (low-dose, high-dose, and positive control groups: 420.80 ± 29.30 pg/ml, 322.33 ± 9.40 pg/ml, 322.97 ± 12.16 pg/ml, respectively), IL-1β (98.69 ± 4.49 pg/ml, 89.02 ± 1.56 pg/ml, 88.88 ± 2.08 pg/ml, respectively), and IL-6 (94.07 ± 3.76 pg/ml, 80.54 ± 3.30 pg/ml, 83.21 ± 3.18 pg/ml, respectively), as well as in their mRNA expression levels (all P < 0.001). In in vitro fibroblast experiments, compared with the control group, the model group exhibited a significant elevation in the supernatant levels of IL-1β (126.42 ± 3.56 pg/ml vs. 34.81 ± 0.44 pg/ml), IL-6 (459.44 ± 9.35 pg/ml vs. 115.51 ± 7.26 pg/ml), CXCL1 (2 434.88 ± 127.63 pg/ml vs. 762.85 ± 30.60 pg/ml) and CXCL12 (3 542.14 ± 35.86 pg/ml vs. 2 095.86 ± 45.12 pg/ml), the expression levels of their mRNAs (all P < 0.001), as well as the protein expression levels of SFRP2, Wnt3a, and β-catenin; after intervention with HLJDT-XJDH-containing serum, all the above indices significantly decreased (all P < 0.001). However, when 20% drug-containing serum intervention was administered simultaneously, the expression of inflammatory factors and chemokines in fibroblasts was significantly higher in the SFRP2 overexpression group than in the non-overexpression group (all P < 0.01). When fibroblasts were co-cultured with HaCaT cells, the model group showed significantly increased cell viability but a decreased apoptosis rate of HaCaT cells compared with the control group, while the low- and high-dose intervention groups showed significantly decreased cell viability but increased apoptosis rates of HaCaT cells compared with the model group (all P < 0.05) . Conclusion:HLJDT-XJDH may exert therapeutic effects in psoriasis by downregulating the SFRP2/Wnt/β-catenin signaling pathway, thereby inhibiting fibroblast activation and inflammatory process, which subsequently suppresses the proliferation of keratinocytes and the activation of inflammatory cells.
9.Comparative study of five physical disability-related standards in China based on ICF and ICD-11
Haiyan YE ; Yaru YANG ; Zhuoying QIU ; Zhongyan WANG ; Di CHEN ; Guiyun SONG ; Fangyong WANG ; Yifan TIAN ; Ye LIU
Chinese Journal of Rehabilitation Theory and Practice 2025;31(7):772-780
Objective To systematically compare five physical disability-related standards widely used in the fields of insurance,work related injury,forensic appraisal,and disability evaluation.The comparison covers definition,coding,classi-fication,grading,and assessment methods to reveal each standard's framework and methodology,and to provide evidence for integrating and optimizing physical disability standards.Methods Based on International Classification of Functioning,Disability and Health(ICF)and International Classifica-tion of Diseases,11th Revision(ICD-11),we analyzed and compared five physical disability-related standards,including Disability Assessment and Code for Life Insurance(Insurance Standard),Grading of Disability due to Human Body Injury(Judicial Standard),Standard for Identify Work Ability—Gradation of Disability Caused by Work-related Injuries and Occupational Diseases(Work Injury Standard),Standard for Assessment of Disability Grades of Military Personnel(Military Standard),and Classification and Grading Criteria of Disability(Disabili-ty Classification Standard).We examined definition,coding systems,classification,grading scales and assess-ment.Results According to the ICF framework,at the level of body functions and structures,physical disability mainly in-volved neuromusculoskeletal and movement related functions(b7),including joint and bone functions(b710-b729),muscle functions(b730-b749),and movement functions(b750-b789).At the activities and participation level,it primarily concerned mobility(d410-d499)and self care(d510-d599).Related conditions were commonly found in Certain infectious or parasitic diseases(01),Diseases of the nervous system(08),Diseases of the muscu-loskeletal system or connective tissue(15),Developmental anomalies(20),and Injury,poisoning or certain other consequences of external causes(22),etc.Among the five standards,only Disability Classification Standard ex-plicitly defined physical disability,yet its terminology did not followed ICF categories.Regarding coding,only the Insurance Standard adopted ICF codes;and the other four used self devised coding systems.The sub catego-ries in these physical disability standards did not fully cover all ICF categories relevant to physical disability.In terms of grading,Disability Classification Standard used a four level grading,whereas the other four standards employed ten level grading.Concerning assessment,all five emphasized evaluation of body structures and func-tions and involved daily activity capacity;however,WHO Disability Assessment Schedule(WHODAS 2.0)for comprehensive functional assessment,had not been proposed for evaluation of general functioning.Conclusion China has developed different physical disability standards to meet diverse needs.There are differences in definition,structure and content.It is necessary to revise and refine the relevant components of physical disabili-ty,including definition,classification,grading,coding and assessment,based on ICF and ICD-11,and to intro-duce WHODAS 2.0 as a comprehensive functional assessment tool.
10.Exploring Pharmacodynamic Material Basis and Mechanism of Hei Xiaoyaosan in Improving Learning and Memory Ability of Insomnia Rats Based on UPLC-Q-TOF-MS and Network Pharmacology
Jiamin LIU ; Hai HUANG ; Pengpeng LIANG ; Yale WANG ; Guiyun LI ; Hongyan WU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(21):19-30
ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), network pharmacology and pharmacodynamics, to investigate the pharmacodynamic material basis and mechanism of Hei Xiaoyaosan in improving learning and memory ability of insomnia rats. MethodUPLC-Q-TOF-MS was used to characterize the chemical constituents of Hei Xiaoyaosan. Network pharmacology was applied to construct the network of active ingredients-intersecting targets-pathways, and molecular docking was performed on key ingredients and core targets. Sixty 8-week-old male SD rats were selected and randomly divided into blank group, model group, Hei Xiaoyaosan low, medium, and high dose groups(3.82, 7.65, 15.30 g·kg-1), and zolpidem tartrate group(0.5 mg·kg-1), with 10 rats in each group. Except for the blank group, the insomnia model was induced by intraperitoneal injection of p-chlorophenylalanine(PCPA) for 4 consecutive days. Rats in each dosing group were administered the corresponding dose by gavage, once a day for 14 consecutive days. Morris water maze test was utilized to assess the learning and memory ability of rats, transmission electron microscopy was employed to examine the ultrastructure of hippocampal synapses, enzyme-linked immunosorbent assay(ELISA) was conducted to analyze the levels of 5-hydroxytryptamine(5-HT), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in hippocampal tissues, and Western blot was performed to detect the expression levels of tumor suppressor protein p53(TP53), rat sarcoma virus(RAS), epidermal growth factor receptor(EGFR), cyclic adenosine monophosphate(cAMP)-response element binding protein(CREB) binding protein(CREBBP), glycogen synthase kinase-3β(GSK-3β), protein kinase B1(Akt1), nitric oxide synthase 1(NOS1), phosphorylated(p)-Akt1, and p-GSK-3β in hippocampal tissues. Additionally, real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was used to assess the mRNA expression levels of TP53, RAS, EGFR, CREBBP, GSK-3β, Akt1 and NOS1. ResultA total of 176 components were identified in Hei Xiaoyaosan, mainly flavonoids, triterpene saponins, phenylpropanoids and other compounds. Network pharmacological analysis revealed that TP53, V-Ha-Ras Harvey Rat sarcoma viral oncogene homolog(HRAS), neuroblastoma sarcoma viral oncogene homolog(NRAS), EGFR, CREBBP, GSK-3β, Akt1 and NOS1 were the key targets of Hei Xiaoyaosan in treating insomnia. The core targets were predominantly associated with cAMP, RAS, Ras-associated protein 1(Rap1), advanced glycation end products(AGE)/receptor for AGE(RAGE), and EGFR signaling pathways, and the key active ingredients of Hei Xiaoyaosan in treating insomnia were 8-shogaol, ligustilide F, 6-gingerol, levistilide A and senkyunolide E. Animal experiment results demonstrated that Hei Xiaoyaosan medium and high dose groups significantly increased body weight, shortened sleep latency and prolonged sleep duration in insomnia rats(P<0.01), significantly decreased escape latency and increased platform crossing frequency(P<0.01), and improved the pathological changes of hippocampal synaptic ultrastructure. Meanwhile, the two groups could significantly elevate 5-HT level, Akt1 mRNA expression, Akt1 and p-Akt1 protein expression(P<0.01), reduce inflammatory factor levels(P<0.01), and down-regulate protein expression levels of TP53, RAS, NOS1, EGFR, CREBBP, GSK-3β and p-GSK-3β(P<0.01), as well as mRNA expression levels of TP53, RAS, NOS1, EGFR, CREBBP and GSK-3β in hippocampal tissues(P<0.01). ConclusionThis study determined that the five key active ingredients(8-shogaol, ligustilide F, 6-gingerol, levistilide A and senkyunolide E) in Hei Xiaoyaosan may improve the learning and memory ability of insomnia rats by regulating signaling pathways such as cAMP, RAS, and EGFR, providing an important reference for its mechanism research and clinical application.


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