1.Luoshi Neiyi Prescription Treats Endometriosis Through TLR4/NF-κB Signaling Pathway
Yuanyuan RUAN ; Sai XU ; Jiangyue TANG ; Xiang LI ; You ZOU ; Fangli PEI ; Lizheng WU ; Kaidi ZHENG ; Shuhong LIN ; Weilan ZHONG ; Cheng ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):185-196
ObjectiveTo investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis (EMs) through the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway. MethodsAnimal experiments were conducted with 50 female SD rats, which were randomized into a sham operation group (10 rats) and a modeling group (40 rats). An autologous endometrial transplantation method was used for the modeling of EMs. The 36 successfully modeled rats were randomly allocated into four groups (n=9 each): The model group, the low-dose (7.87 g·kg-1) Luoshi Neiyi prescription group, the high-dose (15.74 g·kg-1) Luoshi Neiyi prescription group, and the dienogest (0.20 mg·kg-1) group. The physiological status and ectopic lesion volumes of rats in each group were observed. Hematoxylin-eosin (HE) staining was used to observe the morphology of the eutopic endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of inflammatory factors including interleukin-1β (IL-1β) and prostaglandin E2 (PGE2) in the serum of EMs rats. Immunohistochemistry was used to detect the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor A (VEGFA) in the eutopic endometrial tissue. Western blot was adopted to determine the protein levels of TLR4, myeloid differentiation factor 88 (MyD88), phosphorylated nuclear factor-κB (p-NF-κB)/NF-κB, MMP-9, and VEGFA in the eutopic endometrial tissue. In the cell experiments, the cell-counting kit-8 (CCK-8) assay was employed to screen the optimal concentration of Luoshi Neiyi prescription-containing serum, and a scratch assay was performed to assess the migration ability of iheESCs cells. Interventions with Luoshi Neiyi prescription-containing serum, resatorvid (TAK-242, a TLR4 inhibitor), and lipopolysaccharides (LPS, a TLR4 agonist) were conducted, and Western blot was used to detect the expression of proteins related to the TLR4/NF-κB signaling pathway. ResultsThis experiment successfully replicated 36 EMs rat models. Compared with the sham operation group, the model group exhibited visible ectopic lesions on the abdominal wall and an increase in the writhing response score (P<0.01). Furthermore, HE staining revealed the model group exhibited a thickened eutopic epithelium, stromal cell disarrangement, and evident infiltration of inflammatory cells. In addition, the model group showed elevated serum levels of IL-1β and PGE2 (P<0.05, P<0.01), increased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue (P<0.01), and up-regulated protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01). Compared with the model group, all treatment groups exhibited a reduction in the ectopic lesion volume (P<0.01). Furthermore, the writhing response scores were decreased in the high-dose Luoshi Neiyi prescription group and the dienogest group (P<0.05, P<0.01). The pathological state of the ectopic endometrial tissue was alleviated to varying degrees in the treatment groups. Low-dose Luoshi Neiyi prescription reduced serum PGE2 levels, and high-dose Luoshi Neiyi prescription and dienogest decreased serum IL-1β and PGE2 levels in EMs rats (P<0.05, P<0.01). The treatment groups showed decreased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01) and down-regulated protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01). The cell experiments showed that 5%, 10%, and 20% Luoshi Neiyi prescription-containing sera significantly reduced the viability and inhibited the migration of iheESCs. Compared with the control group, 5%, 10%, and 20% Luoshi Neiyi prescription-containing serum groups and the TAK-242 group showed reduced protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA (P<0.01). Compared with the control group, the LPS group showed increased expression of the above proteins (P<0.01). Compared with the LPS group, the LPS+5%, 10%, and 20% Luoshi Neiyi prescription-containing serum groups showed reduced expression of the above proteins (P<0.05, P<0.01). ConclusionLuoshi Neiyi prescription may modulate the TLR4/NF-κB signaling pathway to reduce the inflammatory response and histopathological damage in the eutopic endometrium and suppress the adhesive, invasive, and angiogenic biological processes in the ectopic endometrial tissue, thereby exerting its therapeutic effect on EMs.
2.Application of micro-video tracing training model to emerging infectious diseases knowledge training in emergency department nurses
Binxia LI ; Yuhong ZHANG ; Meiqiong YAN ; Li FENG ; Ping WANG ; Fangli ZHENG ; Jia'nan QIN
Chinese Journal of Medical Education Research 2024;23(1):108-113
Objective:To explore the impact of applying micro-video tracing training model to emerging infectious diseases (EIDs) knowledge training on the autonomous learning ability and learning outcomes among emergency department nurses.Methods:In this study, the training content was developed from the perspective of the historical development of EIDs. The training program was constructed in the form of a micro-video classification module and periodic WeChat group push-based autonomous learning. Convenience sampling was used to select 85 emergency nurses ( n=85) from a grade A tertiary general hospital in Shanghai for micro-video tracing training. The nurses were provided with EIDs knowledge training using a module consisting of 11 episodes, each of which included 5 sessions, i.e., enlightening thinking, micro-video learning, key point consolidation, key point analysis, and extension and expansion. After the intervention, the nurses were compared for their autonomous learning ability and their knowledge, belief, and action levels of EIDs, and their degree of satisfaction with the training model was evaluated. SPSS 20.0 and GraphPad Prism 9.3.0 were used for non-parametric Wilcoxon paired tests of quartiles ( Q25, M, and Q75). Results:After the training, the emergency department nurses had significantly higher scores in overall autonomous learning ability ( Z=-4.69, P<0.001) and in each dimension: concept of self-motivation ( Z=-4.48, P<0.001), task analysis ( Z=-4.06, P<0.001), self-monitoring and regulation ( Z=-4.50, P<0.001), and self-evaluation ( Z=-4.82, P<0.001); they had significantly higher scores in overall knowledge, belief, and action levels of EIDs ( Z=-4.65, P<0.001) and in each dimension: knowledge ( Z=-4.04, P<0.001), belief ( Z=-3.24, P=0.001), and action ( Z=-4.05, P<0.001). Emergency department nurses had a high degree of satisfaction with the training model (degree of satisfaction for each item >90%). Conclusions:The micro-video tracing training model can effectively improve the autonomous learning ability of emergency department nurses and their knowledge, belief, and action levels of EIDs, with a high degree of satisfaction among the nurses, making it promising for wide use.
3.Correlation analysis of self-esteem and imposter phenomenon among clinical nurses
Juan PENG ; Hongyan TAO ; Fangli WEN ; Juan GUI ; Xiaojuan ZHENG ; Yan TANG ; Jieyan WANG
Chinese Journal of Practical Nursing 2023;39(36):2848-2853
Objective:To investigate the correlation relationship between self-esteem and imposter phenomenon among nurses,and to provide reference for optimizing nurse team.Methods:A total of 836 nurses were selected from February to March 2023 in the Yongzhou Central Hospital of Hunan Province and the People′s Hospital of Longhua, Shenzhen. A cross-sectional survey was conducted on nurses using the General Data Questionnaire, Impostor Phenomenon Scale and Self-Esteem Scale.Results:The imposter phenomenon score of nurses was (48.97 ± 12.58) points and the self-esteem score was (28.93 ± 3.86) points. The total score of self-esteem was negatively correlated with the total score of imposter phenomenon ( r= -0.433, P<0.01). Multivariate linear regression analysis showed that with the increase of self-esteem score, the score of imposter phenomenon decreased ( B=-1.402, P<0.01). Self-esteem was an important factor affecting the imposter phenomenon among clinical nurses, accounting for 18.9% of the total variation. Conclusions:The self-esteem and imposter phenomenon of clinical nurses are both at a moderate level, the improvement of self-esteem is beneficial to decreased their imposter phenomenon.
4.Mechanism study on genesis and development of enteritis induced by deoxycholic acid-mediating dysbosis of intestinal microbiota and bile acid abnormal metabolism
Mengsha CEN ; Yubin ZHU ; Yuqin SHEN ; Fangli CHENG ; Xia ZHENG ; Weiling HU ; Ning DAI ; Mengque XU
Chinese Journal of Inflammatory Bowel Diseases 2021;05(1):77-83
Objective:To observe and analyze the influence of deoxycholic acid (DCA) on intestinal microbiota and bile acid metabolism during the process of enteritis induced by DCA in mice.Methods:Twenty C57BL/6J mice were randomly and equally divided into DCA group and control group. The mice in DCA group were developed by feeding the feed with 0.2% DCA and the mice in control group were developed with routine feed for 24 weeks. The degree of intestinal tissue inflammation was evaluated by HE staining and scored, the change of intestinal microbiota was analyzed by pyrosequencing, bile acid levels of all grades in faeces were detected by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) , the expressions of bile acid-related genes were detected by quantitive real-time PCR.Results:The intestinal histology scores in DCA group was significantly higher than that of the control group (both P<0.05) . The diversity of fecal microbiota in the DCA group was significantly reduced, the percentage of Firmicutes at the phylum level and the percentage of Clostridium XIVa at the genus level were significantly decreased (both P<0.05) . The total bile acid, secondary bile acid, unconjugated bile acid and tauro-α-muricholic acid (T-α-MCA) of mice faeces in DCA group were significant higher than those in control group (all P<0.05) . The expression of biliary acid transporter gene organic solute transporter β ( Ost- β) , farnesoid X receptor ( FXR) and fibroblast growth factor 15 ( FGF15) in DCA group decreased significantly (all P<0.05) , while the expressions of liver bile synthesis rate-limiting enzymes Cyp7a1, Cyp7b1 and Cyp27a1 increased obviously (all P<0.05) . Conclusions:DCA can induce the progress of enteritis, which may be related to its destruction of intestinal microbiota in mice and the promotion of liver bile acid synthesis through FXR-FGF15 signaling pathway.
5.Mechanism study on genesis and development of enteritis induced by deoxycholic acid-mediating dysbosis of intestinal microbiota and bile acid abnormal metabolism
Mengsha CEN ; Yubin ZHU ; Yuqin SHEN ; Fangli CHENG ; Xia ZHENG ; Weiling HU ; Ning DAI ; Mengque XU
Chinese Journal of Inflammatory Bowel Diseases 2021;05(1):77-83
Objective:To observe and analyze the influence of deoxycholic acid (DCA) on intestinal microbiota and bile acid metabolism during the process of enteritis induced by DCA in mice.Methods:Twenty C57BL/6J mice were randomly and equally divided into DCA group and control group. The mice in DCA group were developed by feeding the feed with 0.2% DCA and the mice in control group were developed with routine feed for 24 weeks. The degree of intestinal tissue inflammation was evaluated by HE staining and scored, the change of intestinal microbiota was analyzed by pyrosequencing, bile acid levels of all grades in faeces were detected by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) , the expressions of bile acid-related genes were detected by quantitive real-time PCR.Results:The intestinal histology scores in DCA group was significantly higher than that of the control group (both P<0.05) . The diversity of fecal microbiota in the DCA group was significantly reduced, the percentage of Firmicutes at the phylum level and the percentage of Clostridium XIVa at the genus level were significantly decreased (both P<0.05) . The total bile acid, secondary bile acid, unconjugated bile acid and tauro-α-muricholic acid (T-α-MCA) of mice faeces in DCA group were significant higher than those in control group (all P<0.05) . The expression of biliary acid transporter gene organic solute transporter β ( Ost- β) , farnesoid X receptor ( FXR) and fibroblast growth factor 15 ( FGF15) in DCA group decreased significantly (all P<0.05) , while the expressions of liver bile synthesis rate-limiting enzymes Cyp7a1, Cyp7b1 and Cyp27a1 increased obviously (all P<0.05) . Conclusions:DCA can induce the progress of enteritis, which may be related to its destruction of intestinal microbiota in mice and the promotion of liver bile acid synthesis through FXR-FGF15 signaling pathway.
6.Effects of Electroacupuncture at Back?shu and Front?mu Points on Expression of GDNF in Colon Tissue of Functional Constipation Mice
Wei ZHANG ; Ying LI ; Lisha LIU ; Fangli LUO ; Qianhua ZHENG
Journal of Nanjing University of Traditional Chinese Medicine 2015;(5):475-479
ABSTRACTOBJECTIVE To explore the mechanism of electroacupuncture at Back?shu and Front?mu points for functional constipation treatment.METHODS 32 mice were randomly divided into control groupmodel groupelectroacupunctureEA group and EGC group.Compound diphenoxylate were gavaged for modelingand electroacupuncture Tianshu and Dachangshu points30 min each time for a total of 5 times treatment.By light microscopyelectron microscopyimmunohisto-chemistry and in situ hybridizationthe morphology of colonic epithelial structure were observedand the level of GDNF pro-tein and mRNA expression in colon were detected.RESULTS The expression of GDNF protein and mRNA in model group was significantly decreasedP <0.01.Electroacupuncture at back?Shu and front?Mu points improved the expression of GDNF protein and mRNAP <0.05and light microscope and electron microscope showed that acupuncture repaired damaged depi-thelial cellsand then improved the intestinal transmission function.The expression of GDNF protein in EGC group showed no significant difference with the model groupP >0.05but significantly decreasedcompared with EA groupP <0.05.GD-NF mRNA was significantly higher than model groupP <0.05but lower than EA group with no significant differenceP>0.05.The morphology change of colon tissue is more serious.CONCLUSION Electroacupuncture at back?Shu and front? Mu points can improve the expression of GDNF protein and mRNA in EGC cellsrepair the colonic epithelial cellsand then improve the intestinal transmission function.
7.Immunity induced by DNA vaccine of plasmid encoding the rhoptry protein 1 gene combined with the genetic adjuvant of pcIFN-γ against Toxoplasma gondii in mice
Hong GUO ; Guanjin CHEN ; Fangli LU ; Haifeng CHEN ; Huanqin ZHENG
Chinese Medical Journal 2001;114(3):317-320
Objective To construct the eukaryotic expression recombinant plasmid, pcIFN-γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3-rhoptry protein 1 (pc-ROP1) combined with pcIFN-γ against Toxoplasma gondii (T. gondii) infection in mice. Methods A fragment of the IFN-γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion. pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100*!μg, and a booster vaccination was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid or normal saline. At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC-ELISA for the determination of IFN-γ, IL-2 and IL-10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera. Results The recombinant plasmid, pcIFN-γ was constructed. The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN-γ, IL-2 and NO in mice injected with pcROP1 and pcIFN-γ were higher than in those injected with pcROP1 alone. There was no difference in IgG antibody levels between the two groups. Conclusion The genetic adjuvant, pcIFN-γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection.
8.Construction of a recombinant plasmid harbouring the rhoptry protein 1 gene of Toxoplasma gondii and preliminary observations on DNA immunity
Guanjin CHEN ; Hong GUO ; FangLi LÜ ; Huanqin ZHENG
Chinese Medical Journal 2001;114(8):837-840
Objective To observe the immune responses elicited in BALB/c mice by a DNA vaccine. A gene encoding rhoptry protein 1 (ROP1) from Toxoplasma gondii (T. gondii) was cloned into vector pcDNA3. Methods Amplifyied gene fragments coding for ROP1 from the genomic DNA of T.gondii ZS2 were inserted into cloning vector, pUC18, and sub-cloned into pcDNA3. Mice were injected at a dosage of 100?μg recombinant plasmid DNA by intramuscular injection and boosted after 2 weeks. pcDNA3 and normal saline were used as control. 30, 50 and 70 days after the second immunization, NK cell activity, T lymphocyte proliferation and sub-clusters and serum IgG antibody were assayed. Results The specific gene fragment coding for ROP1 was amplified and a pcROP1 recombinant was constructed. At 30 days after immunization, the spleens of the mice were obviously enlarged evidently. NKC activity and the proliferation of spleen T lymphocytes seen on MTT assay were higher in pcROP1 group than in the controls. The number of CD4+ T cells exhibited no obvious increase compared with that of the control, but CD8+ T cells were obviously increased (P<0.05). At 90 days after vaccination, the titer of IgG antibody in the serum of vaccinated mice was positive (1∶100). Conclusion pcROP1 was constructed and it could elicit both cellular and humoral immune responses in immunized mice.

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