1.Determination of gallic acid in sicaokeyin gel by HPLC method
Lianyan YANG ; Weirong XIAO ; Shiyu WEN ; Xinxin XIE ; Hongmin ZHANG ; Bin CHEN
Journal of Pharmaceutical Practice and Service 2026;44(9):468-471
Objective Sanguisorba officinalis is the principal medicinal ingredient of Sicaokeyin gel. Gallic acid is commonly used as a key indicator component for evaluating the quality of Sanguisorba officinalis and its preparations. To establish a HPLC method for the determinant of gallic acid in Sicaokeyin gel. Methods With gallic acid as reference substance,HPLC analysis was performed on a Agilent Eclipse XDB-C18 column (4.6 mm ×25 mm,5 μm) with methanol-0.05%phosphoric acid (5∶95) under isocratic elution. The flow rate was 1.0 ml/min and the detection wavelength was 272 nm. The column temperature was 30℃, the injection volume was 10 μl. Results Gallic acid showed good linear relationship in the range of 1.004-50.18 μg/ml (r=0.999 9). The average recovery was 102.9% (n=9). Conclusion The method was easy to operate; the result was accurate and reliable, which could be used for the quality control of sanguisorba officinalis in Sicaokeyin gel.
2.Chaihu and Longgu Mulitang Regulates ERK/CREB Signaling Pathway to Ameliorate Hippocampal Nerve Injury in Mouse Model of Depression
Shiyu JI ; Li WANG ; Zhuo ZHANG ; Yingzhe GAO ; Zefeng ZHANG ; Siyu CHEN ; Guangjing XIE ; Ping WANG ; Panpan HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):1-9
ObjectiveTo investigate the effects of Chaihu and Longgu Mulitang (CLMT) on hippocampal neural damage in the mouse model of depression via the extracellular signal-regulated protein kinase (ERK)/cAMP-response element-binding protein (CREB) signaling pathway. MethodsSeventy-eight male C57BL/6 mice were randomly allocated into normal control, model, low/medium/high-dose (2.89, 5.78, and 11.56 g·kg-1, respectively) CLMT, and paroxetine (10 mg·kg-1) groups. A depression model was established by chronic unpredictable mild stress (CUMS) combined with social isolation. Behavioral tests were carried out to evaluate depressive-like behaviors. Hematoxylin-eosin staining and Nissl staining were performed to assess hippocampal morphology and neuronal damage. Immunofluorescence was employed to detect glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba1). Real-time PCR was employed to measure the mRNA levels of ERK and CREB. Western blot was employed to determine the expression of ERK/CREB pathway proteins and brain-derived neurotrophic factor (BDNF) in the hippocampal tissue. Molecular Operating Environment (MOE) software was used for molecular docking to evaluate the interactions between CLMT components and target proteins. ResultsCompared with the normal control group, the model group showed decreased sucrose preference (P0.01), increased tail-suspension immobility time (P0.01), decreased activity in the central region of the open field test (P0.01), and decreased activity in the middle and open-arm region of the elevated plus maze test (P0.01). The hippocampal area in the model group showed wrinkled cells and a reduction in the number of cells, neurons with reduced sizes and Nissl bodies, enhanced fluorescence intensity of GFAP and Iba1 (P0.01), and down-regulated expression of phosphorylated (p)-ERK, p-CREB, and BDNF (P0.05, P0.01) and mRNA levels of ERK and CREB (P0.01). Compared with the model group, the CLMT group showed increased body weight (P0.05, P0.01), restored cell morphology, with only a small number of ruptured cells, normal neuronal structure and morphology with obvious nuclei and abundant Nissl bodies, weakened fluorescence intensity of GFAP and Iba1 (P0.05, P0.01), up-regulated mRNA levels of ERK and CREB (P0.05, P0.01) and protein levels of phosphorylated (p)-ERK, p-CREB, and BDNF in the hippocampal tissue (P0.05, P0.01). The results of molecular docking indicated that nine active ingredients in CLMT had good binding affinity with ERK and CREB. ConclusionCLMT may ameliorate the hippocampal nerve injury in the mouse model of depression by regulating the ERK/CREB pathway.
3.Smad3 signaling pathway promotes silica-induced epithelial-mesenchymal transition
Shiyu YANG ; Yingdie ZHANG ; Yujia XIE ; Haoyu YIN ; Pei GU ; Dongming WANG ; Weihong CHEN ; Jixuan MA
Chinese Journal of Industrial Hygiene and Occupational Diseases 2025;43(4):241-246
Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.
4.NMRAL1 promotes proliferation and migration of papillary thyroid carcinoma cells
Yiting XIE ; Xinyi LONG ; Shiyu CAO ; Lin XIAO ; Tengyun MA ; Feng YE
Chinese Journal of Clinical and Experimental Pathology 2025;41(1):14-22
Purpose To explore the role of translesion synthesis-related gene NmrA like redox sensor 1(NMRAL1)in the development of papillary thyroid carcinoma(PTC).Methods Bioinformatics analysis was em-ployed to investigate the expression of the NMRAL1 gene across various cancer types and its correlation with prognosis.Immunohistochemistry was utilized to analyze the expression pattern of NMRAL1 in PTC and conduct clinicopathological correlation analysis.Additionally,stable cell lines overexpressing NMRAL1 were established in TPC-1 cells.The effects of NMRAL1 overexpression on cell proliferation,migration,and invasion of TPC-1 and Nthy-ori 3-1 cells were assessed using CCK-8 assay,clonogenic assay,Transwell assay,and scratch assay,respectively.Furthermore,RT-qPCR experiments were conducted to investigate the regulatory effect of NMRAL1 on the expression of genes associated with translesion DNA synthesis.Results Bioinformatics analysis revealed that NMRAL1 was highly expressed in all 26 types of tumors compared to normal tissues in the TCGA x GTEx dataset.Patients with high expression of NMRAL1 in thyroid cancer had significantly lower survival rates than those with low expression(P<0.05).In the immunohisto-chemical validation of 30 PTC samples,NMRAL1 had a positive rate of 73.33%,in PTC tissues,while it was not ex-pressed in adjacent normal thyroid tissues.The staining intensity scores of NMRAL1 in PTC and adjacent tissues exhib-it significant differences(P<0.000 1).High expression of NMRAL1 was associated with the size of the tumor(P=0.027 4)and lymph node metastasis(P=0.044 1).In vitro functional experiments revealed a significant enhance-ment in both cell proliferation and migration/invasion capacities upon overexpression of NMRAL1(P<0.05).Moreo-ver,mRNA and protein expression levels of translesion synthesis-related genes were upregulated(P<0.001).Con-clusion NMRAL1 promotes proliferation and migration of thyroid carcinoma cells,suggesting that this function may be achieved by enhancing DNA damage repair capacity,thereby improving the survival of damaged cells.
5.NMRAL1 promotes proliferation and migration of papillary thyroid carcinoma cells
Yiting XIE ; Xinyi LONG ; Shiyu CAO ; Lin XIAO ; Tengyun MA ; Feng YE
Chinese Journal of Clinical and Experimental Pathology 2025;41(1):14-22
Purpose To explore the role of translesion synthesis-related gene NmrA like redox sensor 1(NMRAL1)in the development of papillary thyroid carcinoma(PTC).Methods Bioinformatics analysis was em-ployed to investigate the expression of the NMRAL1 gene across various cancer types and its correlation with prognosis.Immunohistochemistry was utilized to analyze the expression pattern of NMRAL1 in PTC and conduct clinicopathological correlation analysis.Additionally,stable cell lines overexpressing NMRAL1 were established in TPC-1 cells.The effects of NMRAL1 overexpression on cell proliferation,migration,and invasion of TPC-1 and Nthy-ori 3-1 cells were assessed using CCK-8 assay,clonogenic assay,Transwell assay,and scratch assay,respectively.Furthermore,RT-qPCR experiments were conducted to investigate the regulatory effect of NMRAL1 on the expression of genes associated with translesion DNA synthesis.Results Bioinformatics analysis revealed that NMRAL1 was highly expressed in all 26 types of tumors compared to normal tissues in the TCGA x GTEx dataset.Patients with high expression of NMRAL1 in thyroid cancer had significantly lower survival rates than those with low expression(P<0.05).In the immunohisto-chemical validation of 30 PTC samples,NMRAL1 had a positive rate of 73.33%,in PTC tissues,while it was not ex-pressed in adjacent normal thyroid tissues.The staining intensity scores of NMRAL1 in PTC and adjacent tissues exhib-it significant differences(P<0.000 1).High expression of NMRAL1 was associated with the size of the tumor(P=0.027 4)and lymph node metastasis(P=0.044 1).In vitro functional experiments revealed a significant enhance-ment in both cell proliferation and migration/invasion capacities upon overexpression of NMRAL1(P<0.05).Moreo-ver,mRNA and protein expression levels of translesion synthesis-related genes were upregulated(P<0.001).Con-clusion NMRAL1 promotes proliferation and migration of thyroid carcinoma cells,suggesting that this function may be achieved by enhancing DNA damage repair capacity,thereby improving the survival of damaged cells.
6.Smad3 signaling pathway promotes silica-induced epithelial-mesenchymal transition
Shiyu YANG ; Yingdie ZHANG ; Yujia XIE ; Haoyu YIN ; Pei GU ; Dongming WANG ; Weihong CHEN ; Jixuan MA
Chinese Journal of Industrial Hygiene and Occupational Diseases 2025;43(4):241-246
Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.
7.Simultaneous Determination of Five Nucleoside Components in Periplaneta Americana by Quantitative Analysis of Multi-components by Single Marker Method
Muyuan CHEN ; Siqi FENG ; Zhenwen QIU ; Qian LUO ; Chunzhu XIE ; Shiyu QI ; Yuechun HUANG ; Huibiao LI
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(7):1055-1060
Objective A HPLC-quantitative analysis of multi-components by single marker(QAMS)was established to determine 5 ingredients including uracil,uridine,hypoxanthine,inosine and guanosine in Periplaneta americana.Methods Separation took place on a Agilent ZORBAX SB-Aq column(250 mm×4.6 mm,5 μm)by gradient elution of methanol-0.01 mol·L-1 potassium dihydrogen phosphate at 20℃with a flow rate of 1.0 mL·min-1.The detection wavelength was 260 nm and the injection amount was 10 μL.The relative correction factors(fa/b)was calculated for the other four components with uridine as an internal standard.The content of 5 ingredients in 10 batches of Periplaneta americana was determined by QAMS.Results were compared with those of external standard method(ESM).Results Five nucleosides showed good linear relationships in their own ranges(r>0.999 5),and the average recoveries ranged from 97.0%to 100.8%.The relative correction factors of uracil,hypoxanthine,inosine and guanosine were 0.908 0,1.005 3,1.969 5 and 1.303 4,respectively.Conclusion The established method is accurate and stable.It can provide theoretical reference for the quality control of Periplaneta americana.
8.Denatonium benzoate promotes MrgprB2-mediated rat mast cell degranulation
Huaping XU ; Xiaoyun SHI ; Jiexin ZOU ; Xin LI ; Mengting XIE ; Shiyu XIAO ; Linbo SHI
Chinese Journal of Immunology 2024;40(10):2037-2041
Objective:To explore the potent effects of denatonium benzoate(DB)on Mas-related G protein-coupled receptor-B2(MrgprB2)-mediated rat mast cell degranulation.Methods:RBL-2H3 cells were treated with DB overnight,before challenged with MrgprB2 ligands substance P(SP).The release of β-hex from MrgprB2-activated RBL-2H3 was detected by substrate method.Detec-tion of LTC4,IL-6,TNF-α and cPLA2 activity were performed by ELISA.The Ca2+influx and the expression of RBL-2H3 MrgprB2 re-ceptors were measured by fluorescence assay.Results:The results showed 10 μmol/L,50 μmol/L,80 μmol/L,100 μmol/L DB treat-ments promoted β-hex and LTC4 releases from activated RBL-2H3,accompanied by increased Ca2+mobilization and cPLA2 activa-tion.DB treatments did not affect IL-6 and TNF-α LTC4 releases in MrgprB2-activated RBL-2H3,as well as the levels of MrgprB2 ex-pression in mast cells.Conclusion:Taken together,DB enhanced the MrgprB2-mediated RBL-2H3 degranulation in vitro,probably by up-regulating Ca2+mobilization in activated cells.
9.Research on the optimization of pharmaceutical care in Chinese social pharmacy based on Kano theory
Yanqi MAO ; Shiyu XIE ; Kan TIAN
China Pharmacy 2023;34(2):133-138
OBJECTIVE To identify the demand levels and specific connotations of pharmaceutical care in social pharmacy based on Kano theory, and to provide suggestions for the optimization of pharmaceutical care in Chinese social pharmacy. METHODS Using Kano theory as the analysis framework, the needs of consumer for different levels of pharmaceutical care in social pharmacy were identified through literature combing. The ideas and suggestions were proposed for the optimization of pharmaceutical care in Chinese social pharmacy based on the content and characteristics of different levels of needs. RESULTS & CONCLUSIONS The demands for pharmaceutical care in Chinese social pharmacy were divided into three levels, among which the basic demand included ensuring the accessibility, safety and effectiveness of drugs; the expectation demand included personalized medication guidance and management, convenient and efficient medication purchasing services triggered by consumer upgrading; the charming demand included health services and management, professional and high-quality service experience. Social pharmacies should take drug security as the core, achieve high quality and good price, and fully meet basic demand; take patient medication management as the grip, conduct double-drive professional services and model innovation to fully respond to expectation demand; take public health as the goal, broaden service content and experience value, and meet the charming demand of consumer at the right time.
10.Observation on the efficacy of different stents in the treatment of patients with advanced esophageal cancer
Shiyu JI ; Mingxin ZHANG ; Huahong XIE ; Yuan BAI ; Tong WANG
Journal of International Oncology 2023;50(2):76-81
Objective:To explore the efficacy and safety of treating advanced esophageal cancer by implanting the common stent and the radioactive 125I particle stent with endoscope. Methods:The clinical data of patients with advanced esophageal cancer admitted to Jingbian County People's Hospital of Shaanxi Province, the First Affiliated Hospital of Xi'an Medical University, Xijing Hospital of Digestive Diseases of Air Force Medical University and the First Hospital of Yulin of Shaanxi Province from December 2014 to December 2020 were retrospectively analyzed. Patients were divided into common stent group ( n=66) and radioactive particle stent group ( n=34) according to different stent types. The postoperative complications, Karnofsky performance status (KPS) score, dysphagia score, restenosis rate and quality of life were compared between the two groups. Results:The incidences of postoperative retrosternal pain in the common stent group and the radioactive particle stent group were 65.2% (43/66) and 47.1% (16/34) respectively. The incidences of pharyngeal pain and hoarseness were 12.1% (8/66) and 5.9% (2/34) . The incidences of abdominal pain were 9.1% (6/66) and 2.9% (1/34) . The incidences of errhysis were 3.0% (2/66) and 2.9% (1/34) . The incidences of vomiting and nausea were 7.6% (5/66) and 5.9% (2/34) respectively. There were no statistically significant differences between the two groups ( χ2=3.04, P=0.081; χ2=0.40, P=0.527; χ2=0.53, P=0.467; χ2<0.01, P>0.999; χ2<0.01, P>0.999) . In the two groups, KPS scores in the first, second, third and sixth month after operation were higher than those before operation (all P<0.05) . KPS scores of the radioactive particle stent group in the second, third and sixth month were significantly higher than those of the common stent group [ (89.73±7.84) points vs. (82.37±7.42) points, t=4.62, P<0.001; (93.63±8.13) points vs. (88.33±7.28) points, t=3.74, P<0.001; (92.78±6.26) points vs. (87.28±8.73) points, t=3.77, P<0.001]. The dysphagia scores of patients in the two groups in the first, second, third and sixth month were lower than those before operation (all P<0.05) . The dysphagia scores of the radioactive particle stent group in the third and sixth month after operation were significantly lower than those of the common stent group [ (0.68±0.12) points vs. (2.33±0.32) points, t=26.20, P<0.001; (0.82±0.22) points vs. (2.67±0.24) points, t=36.92, P<0.001]. In the third month after operation, the restenosis rate of the radioactive particle stent group was significantly lower than that of the common stent group [5.88% (2/34) vs. 42.4% (28/66) , χ2 =14.27, P<0.001]. The scores of QLQ-C30 and OES-18 scales in the first, second, third and sixth month after operation were lower than those before operation (all P<0.05) . The scores of QLQ-30 scale in the radioactive particle stent group in the second, third and sixth month were significantly lower than those in the common stent group [ (19.12±3.02) points vs. (21.22±2.87) points, t=3.39, P=0.001; (15.04±1.68) points vs. (20.43±2.23) points, t=12.39, P<0.001; (14.38±2.18) points vs. (19.77±3.67) points, t=9.20, P<0.001]. The scores of OES-18 scale in the radioactive particle stent group were also significantly lower than those in the common stent group [ (17.13±2.07) points vs. (20.64±2.11) points, t=7.95, P<0.001; (15.22±1.88) points vs. (19.24±1.76) points, t=10.62, P<0.001; (14.74±2.36) points vs. (18.53±3.27) points, t=6.01, P<0.001]. Conclusion:The radioactive particle stent can improve the quality of life of patients with advanced esophageal cancer with esophageal stenosis, so as to improve dysphagia and reduce the restenosis rate after operation. However, whether it is obviously superior to common stent in prolonging survival time and reducing complications needs to be further confirmed by a multicenter, prospective, large-sample randomized controlled study.

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