1.Mechanisms of Huanglian Jiedutang and Its Major Active Constituents in Inhibiting LPS-induced M1 Polarisation of BV2 Microglia
Haojia ZHANG ; Kai WANG ; Kunjing LIU ; Xin LAN ; Zijin SUN ; Chunyu WANG ; Wenyuan MA ; Wei SHAO ; Jinhua HAN ; Liyang DONG ; Changxiang LI ; Xueqian WANG ; Youxiang CUI ; Fafeng CHENG ; Qingguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):44-55
ObjectiveTo investigate whether Huanglian Jiedutang (HLJD) and its major active constituents (geniposide, baicalin, and berberine) can inhibit the inflammatory response of BV2 cells under lipopolysaccharide (LPS) stimulation via the high-mobility group protein B1 (HMGB1)/Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway, and to explore differences in therapeutic efficacy among the three monomers, their combined formula, and HLJD under equal content ratios. MethodsBV2 microglial cells were used as the primary experimental model. Cell viability was assessed using the cell counting kit-8 (CCK-8) method to examine the effects of different concentrations of dimethyl sulfoxide (DMSO, 0.8%, 0.4%, 0.2%, 0.1%, and 0.05%) on cell viability. IncuCyte was employed to monitor the growth of cells under different concentrations of HLJD (200, 100, 50, 25, 12.5, 6.25 mg·L-1). Nitric oxide (NO) assay was used to screen the optimal HLJD concentration. High-performance liquid chromatography (HPLC) determined the content of geniposide, baicalin, and berberine in HLJD, and experimental groups were subsequently established according to the relative proportions of these constituents. CCK-8 assay evaluated cell viability under different treatments. Enzyme-linked immunosorbent assay (ELISA) measured levels of inflammatory factors (TNF-α, IL-1β, IL-6, IL-10) in the supernatant. Flow cytometry assessed the effects of treatments on M1-type polarization of BV2 cells. Western blot determined the expression levels of HMGB1, TLR4, and NF-κB-related proteins. ResultsCompared with the blank group, DMSO at concentrations ≤0.2% did not affect cell viability within 48 h. BV2 cell growth plateaued at 24 h after treatment with 200 mg·L-1 HLJD. Under stimulation with 2 mg·L-1 LPS, this concentration of HLJD effectively reduced NO release, and 6 h pre-treatment had a stronger inhibitory effect on NO than direct administration. HPLC results showed that 1 mg of HLJD freeze-dried powder contained approximately 24 μg of geniposide, 15 μg of baicalin, and 30 μg of berberine. Based on these ratios, experimental groups were blank, LPS (2 mg·L-1), HLJD (200 mg·L-1), monomer combination, geniposide (4.8 mg·L-1), baicalin (3 mg·L-1), and berberine (6 mg·L-1). The monomer combination group consisted of all three active constituents dissolved together. LPS and HLJD or its active constituents did not affect cell viability compared with the blank group. LPS significantly increased TNF-α, IL-1β, IL-6, and IL-10 in the supernatant (P<0.01). HLJD and its active constituents significantly reduced pro-inflammatory factors TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01) while upregulating anti-inflammatory IL-10 (P<0.01), with the monomer combination showing the strongest effect (P<0.05, P<0.01). Compared with the blank group, LPS significantly increased the proportion of CD80⁺CD86⁺ (M1-type) BV2 cells (P<0.01). HLJD and its constituents partially inhibited M1 polarization (P<0.05, P<0.01), with the monomer combination exhibiting the most pronounced effect (P<0.05, P<0.01). Compared with the blank group, LPS upregulated HMGB1, TLR4, and NF-κB-related proteins (P<0.01), whereas HLJD and its active constituents significantly reduced their expression (P<0.05, P<0.01), with the monomer combination having the strongest regulatory effect (P<0.05, P<0.01). ConclusionHLJD and its major active constituents (geniposide, baicalin, berberine) can inhibit LPS-induced inflammatory responses in BV2 cells. The combination of the three active constituents demonstrates the most potent anti-inflammatory effect, significantly attenuating M1-type polarization of BV2 cells via the HMGB1/TLR4/NF-κB signaling pathway.
2.Rapid enrichment and SERS differentiation of various bacteria in skin interstitial fluid by 4-MPBA-AuNPs-functionalized hydrogel microneedles.
Ying YANG ; Xingyu WANG ; Yexin HU ; Zhongyao LIU ; Xiao MA ; Feng FENG ; Feng ZHENG ; Xinlin GUO ; Wenyuan LIU ; Wenting LIAO ; Lingfei HAN
Journal of Pharmaceutical Analysis 2025;15(3):101152-101152
Bacterial infection is a major threat to global public health, and can cause serious diseases such as bacterial skin infection and foodborne diseases. It is essential to develop a new method to rapidly diagnose clinical multiple bacterial infections and monitor food microbial contamination in production sites in real-time. In this work, we developed a 4-mercaptophenylboronic acid gold nanoparticles (4-MPBA-AuNPs)-functionalized hydrogel microneedle (MPBA-H-MN) for bacteria detection in skin interstitial fluid. MPBA-H-MN could conveniently capture and enrich a variety of bacteria within 5 min. Surface enhanced Raman spectroscopy (SERS) detection was then performed and combined with machine learning technology to distinguish and identify a variety of bacteria. Overall, the capture efficiency of this method exceeded 50%. In the concentration range of 1 × 107 to 1 × 1010 colony-forming units/mL (CFU/mL), the corresponding SERS intensity showed a certain linear relationship with the bacterial concentration. Using random forest (RF)-based machine learning, bacteria were effectively distinguished with an accuracy of 97.87%. In addition, the harmless disposal of used MNs by photothermal ablation was convenient, environmentally friendly, and inexpensive. This technique provided a potential method for rapid and real-time diagnosis of multiple clinical bacterial infections and for monitoring microbial contamination of food in production sites.
3.Discovery of toad-derived peptide analogue targeting ARF6 to induce immunogenic cell death for immunotherapy of hepatocellular carcinoma.
Dihui XU ; Xiang LV ; Meng YU ; Ao TAN ; Jiaojiao WANG ; Xinyi TANG ; Mengyuan LI ; Wenyuan WU ; Yuyu ZHU ; Jing ZHOU ; Hongyue MA
Journal of Pharmaceutical Analysis 2025;15(3):101038-101038
Image 1.
4.Rapid enrichment and SERS differentiation of various bacteria in skin interstitial fluid by 4-MPBA-AuNPs-functionalized hydrogel microneedles
Ying YANG ; Xingyu WANG ; Yexin HU ; Zhongyao LIU ; Xiao MA ; Feng FENG ; Feng ZHENG ; Xinlin GUO ; Wenyuan LIU ; Wenting LIAO ; Lingfei HAN
Journal of Pharmaceutical Analysis 2025;15(3):564-576
Bacterial infection is a major threat to global public health,and can cause serious diseases such as bacterial skin infection and foodborne diseases.It is essential to develop a new method to rapidly di-agnose clinical multiple bacterial infections and monitor food microbial contamination in production sites in real-time.In this work,we developed a 4-mercaptophenylboronic acid gold nanoparticles(4-MPBA-AuNPs)-functionalized hydrogel microneedle(MPBA-H-MN)for bacteria detection in skin inter-stitial fluid.MPBA-H-MN could conveniently capture and enrich a variety of bacteria within 5 min.Surface enhanced Raman spectroscopy(SERS)detection was then performed and combined with ma-chine learning technology to distinguish and identify a variety of bacteria.Overall,the capture efficiency of this method exceeded 50%.In the concentration range of 1 × 10 7 to 1 × 10 10 colony-forming units/mL(CFU/mL),the corresponding SERS intensity showed a certain linear relationship with the bacterial concentration.Using random forest(RF)-based machine learning,bacteria were effectively distinguished with an accuracy of 97.87%.In addition,the harmless disposal of used MNs by photothermal ablation was convenient,environmentally friendly,and inexpensive.This technique provided a potential method for rapid and real-time diagnosis of multiple clinical bacterial infections and for monitoring microbial contamination of food in production sites.
5.A nomogram risk prediction model for symptomatic cerebrovascular ischaemia based on carotid intra-plaque neovascularisation
Wenyuan MA ; Qi XU ; Yamei MA ; Yinghui CHEN
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(11):1310-1314
Objective To construct a nomogram risk prediction model for symptomatic cerebrovas-cular ischaemia based on intra-plaque neovascularisation in carotid arteries in patients with ische-mic cerebrovascular disease(ICVD).Methods A retrospective study was conducted on 320 ICVD patients who were consecutively admitted to Wuhan Fourth Hospital from April 2020 to April 2024.In a ratio of 3∶1,240 cases were assigned into a training set and 80 cases into a validation set.The patients in the training set were further divided into 147 cases in the symptomatic sub-group and 93 cases in the asymptomatic subgroup according to the presence or absence of relevant symptoms or signs.Multifactorial logistic regression analysis was used to identify the risk factors for symptomatic ischemia in ICVD patients,and a nomogram risk prediction model was construc-ted and the prediction efficacy of the model was evaluated.Results The proportions of plaque multiplicity,ulcerated plaque,stenosis ≥70%,and intra-plaque neovascularization were signifi-cantly higher in the symptomatic subgroup than the asymptomatic subgroup(P<0.05,P<0.01).Multifactorial logistic regression analysis showed that plaque multiplicity(OR=1.261,95%CI:1.088-1.539,P=0.003),ulcerated plaque(OR=1.458,95%CI:1.132-1.661,P=0.001),and stenosis ≥70%(OR=2.023,95%CI:1.458-2.561,P=0.001),and intra-plaque neovasculariza-tion(OR=1.206,95%CI:1.057-1.489,P=0.002)were independent risk factors for the occur-rence of symptomatic cerebral ischemia in ICVD patients.H-L deviation test showed that the con-structed nomogram risk prediction model had a good fit(x2=9.362,P=0.295).Internal and ex-ternal validation showed that the calibration curves for both the training and validation sets were consistent with the original curves,and the AUC value was were 0.871 and 0.864,respectively.De-cision curve analysis showed that the model had a significant standardized clinical net benefit when the predicted risk threshold exceeded 0.01.Conclusion Ultrasonography is instructive in as-sessment of the presence or absence of intra-plaque neovascularization in carotid atherosclerotic plaques.And our constructed nomogram risk prediction model has good predictive value for the development of symptomatic cerebral ischemia.
6.Immunological mechanisms in steatotic liver diseases: An overview and clinical perspectives
Mengyao YAN ; Shuli MAN ; Long MA ; Lanping GUO ; Luqi HUANG ; Wenyuan GAO
Clinical and Molecular Hepatology 2024;30(4):620-648
Steatotic liver diseases (SLD) are the principal worldwide cause of cirrhosis and end-stage liver cancer, affecting nearly a quarter of the global population. SLD includes metabolic dysfunction-associated alcoholic liver disease (MetALD) and metabolic dysfunction-associated steatotic liver disease (MASLD), resulting in asymptomatic liver steatosis, fibrosis, cirrhosis and associated complications. The immune processes include gut dysbiosis, adiposeliver organ crosstalk, hepatocyte death and immune cell-mediated inflammatory processes. Notably, various immune cells such as B cells, plasma cells, dendritic cells, conventional CD4+ and CD8+ T cells, innate-like T cells, platelets, neutrophils and macrophages play vital roles in the development of MetALD and MASLD. Immunological modulations targeting hepatocyte death, inflammatory reactions and gut microbiome include N-acetylcysteine, selonsertib, F-652, prednisone, pentoxifylline, anakinra, JKB-121, HA35, obeticholic acid, probiotics, prebiotics, antibiotics and fecal microbiota transplantation. Understanding the immunological mechanisms underlying SLD is crucial for advancing clinical therapeutic strategies.
7.Construction of the evaluation index system for core competence of hospital specialist service operation assistants
Min WANG ; Xiao ZHANG ; Dawei QIN ; Meirong LYU ; Cong SHI ; Xingyan MEI ; Tiantian WU ; Wenyuan MA
Chinese Journal of Hospital Administration 2023;39(9):692-697
Objective:To construct an evaluation index system for the core competence of hospital specialist service operation assistants and provide reference for the evaluation of such competence.Methods:From January to March 2022, literature analysis and behavioral event interviews were used to initially establish a core competence evaluation index system of hospital specialist service operation assistants, based on the Donabedian model. Subsequently, the Delphi expert consultation method was applied to conduct correspondence consultation, inviting experts to evaluate the contents and importance of the index system, using analytic hierarchy process to determine the weights of the indexes at all levels.Results:Two rounds of expert consultation were carried out, and the valid recovery rate of the questionnaire was 100%. The familiarity coefficient of the second round of correspondence was 0.87, the basis of judgment coefficient was 0.90, and the authority coefficient was 0.89. The final evaluation index system for core competence of hospital specialist service operation assistant consisted of 3 first-level indexes, 13 second-level indexes and 81 third-level indexes. The weight of the first-level index structure index was 0.266, and the highest weight among the second-level indexes was the operational development ability (0.083), while the highest weight among the third-level indexes of operational development ability was the comprehensive coordination ability (0.193); The weight of the first-level index process index was 0.405, and the corresponding second-level and third-level indexes with the highest weight were department operation practice work (0.157) and reasonable resource allocation (0.303), respectively; The weight of the first-level index result index was 0.329, and the corresponding second-level and third-level indexes with the highest weight were the weight of medical quality and safety (0.103) and drug adverse reaction reporting rate (0.237), respectively.Conclusions:The evaluation index system constructed in this study proves scientific and reasonable in weight assignment, proving a reference for the management of the specialist service operation assistants.
8.Association between early life exposure to famine and risk for subtype and classification of hypertension in middle and old age
Chenxi LI ; Wenyuan MA ; Zhiyu LIU ; Yaojia SHEN ; Xinxin YE ; Qian YI ; Peige SONG
Journal of Public Health and Preventive Medicine 2023;34(3):1-6
Objective To investigate the relationship between exposure to famine in early life stage and hypertension phenotype and grade in middle and old age. Methods People born between 1951 and 1965 in the 2015 China Health and Elderly Care Follow-up Survey were included in the study, and were divided into unexposed group, fetal exposed group, childhood exposed group and adolescent exposed group according to the time of famine occurrence and birth year of the participants. Logistic regression model was used to explore the effects of different famine exposure periods in early life stage on hypertension classification (including normal high value, grade I, grade II and grade III) and phenotype (including isolated systolic hypertension[ISH], isolated diastolic hypertension [IDH] and combined systolic and diastolic hypertension [SDH]). Results Compared with unexposed group, fetal famine exposure (OR=1.59, 95% CI :1.10-2.30), childhood famine exposure (OR=1.67, 95% CI :1.04-2.70) and adolescent famine exposure (OR=3.42, 95% CI : 2.51-4.66) were the risk factors for ISH. Only famine exposure during adolescence (OR=1.54, 95% CI: 1.07-2.21) was a risk factor for SDH. In addition, fetal famine exposure (OR=1.41, 95% CI: 1.05-1.89) and adolescent famine exposure (OR=2.22 , 95% CI: 1.71-2.88) were risk factors for developing grade I hypertension. Famine exposure in childhood (OR=2.45, 95% CI: 1.21-4.94) and famine exposure in adolescence (OR=2.45, 95% CI: 1.44-4.19) were risk factors for grade 2 hypertension. Conclusion Famine exposure in early life stage was associated with the phenotype and grade of hypertension. Therefore, balanced nutrition in early life is important to prevent hypertension in adulthood.
9.Air quality of metro system in a northwestern city of China from 2021 to 2022
Ying ZHANG ; Yuxi MA ; Yong ZHANG ; ABULIMITI ABABAIKELI ; Wenyuan HAO ; Haofeng YANG
Journal of Environmental and Occupational Medicine 2023;40(11):1290-1296
Background Most metro system are underground, airtight, with inadequate ventilation and massive gatherings, posing health risks to metro riders. Objective To evaluate air quality of Metro Line 1 in a city, and provide suggestions and basis for preventing harmful factors and protecting the health of passengers. Methods Station halls, station platforms, and metro carriages of Metro Line 1 in a city were monitored in summer (from July to August in 2021) and winter (from January to February in 2022). Six metro stations were selected by stratified sampling. Each station and carriage were monitored for three consecutive days in rush hours (9:00–11:00 and 19:00–21:00) and non-rush hours (11:00–13:00), with the same monitoring frequency. The monitored indicators were physical factors (temperature, relative humidity, wind speed, illumination, and noise), chemical factors (carbon monoxide, carbon dioxide, inhalable particles, formaldehyde, benzene, toluene, xylene, ammonia, and ozone), biological factor (airborne total bacterial count), and radiation factor (radon). The monitoring results were compared by location, time period, and season. Results According to the Hygienic indicators and limits for public places (GB 37488—2019), the selected physical factors did not meet the standard, especially the temperature and relative humidity of station hall and platform, and wind speed and noise of carriage. The results of physical factors varied significantly by location (P<0.05). In summer, the temperature of carriage [M (P25, P75), 23.9 (23.3, 24.6)℃] was the lowest, and the wind speed [0.78 (0.37, 1.11) m·s−1] and noise [76.0 (72.0, 80.3) dB] of carriage were the highest; in winter, the temperatures of station hall and platform were the lowest [16.2 (13.2, 17.2)℃ and 16.2 (13.4, 17.0)℃, respectively], the relative humidity of carriage [26.4% (24.2%, 27.9%)] was the lowest, and the wind speed and noise of carriage were the highest [0.83 (0.47, 1.18) m·s−1 and 74.5 (70.1, 78.3) dB, respectively]. The physical factors varied significantly by time period (P<0.05). In summer, the temperature was the lowest during morning rush hours [24.0 (23.0, 24.8)℃] and non-rush hours [24.2 (23.2, 24.9)℃], and the relative humidity during evening rush hours was the lowest [41.9% (37.0%, 47.8%)]; in winter, the temperature was the lowest during morning and evening rush hours [16.8 (13.4, 19.7)℃ and 16.5 (15.1, 19.4)℃, respectively], the relative humidity during the non-rush period was the lowest [26.8% (24.7%, 28.6%)], and the wind speed during evening rush hours was the highest [0.28 (0.19, 0.51) m·s−1]. All measured chemical factors, biological factor, and radiation factor met the national standard (GB 37488—2019). Conclusion The chemical, biological, and radiative factors are complied with the national standard (GB 37488—2019) except physical factors such as temperature, relative humidity, wind speed, and noise. We suggest that the metro operators make full use of air conditioning system in combination with humidifiers to better regulate temperature and relative humidity, and .arrange working hours reasonably and provide noise-proof earplugs for carriage staff to protect against noise hazard.
10.Determination of Free Lanolin Alcohol and Residual Pesticides in Lanolin
DING Yinmeng ; LIU Jing ; MA Qingxiu ; LIU Wenyuan
Chinese Journal of Modern Applied Pharmacy 2023;40(14):1976-1982
OBJECTIVE To determine the content of free lanolin alcohol and residual pesticides in lanolin by gas chromatography, and to evaluate the feasibility of the established method. METHODS Gel permeation chromatography(GPC) was used for pretreatment, and the contents of free lanolin alcohol and residual pesticides were determined by GC-FID and GC-MS, respectively. And the qualitative analysis was conducted based on the retention time and quantitative analysis was conducted by total peak area. RESULTS The recovery of GPC pretreatment system met the requirements. Under the two established methods, the determinand were well separated; the satisfactory linearity with good correlation coefficients >0.999 5 were obtained; and the average recoveries were in the range of 90.49%-102.78%(n=6). The content of free lanolin alcohol in 3 batches of lanolin was <2.84%, cis-permethrin and trans-permethrin were determined in lanolin and the other residual pesticides were not detected. CONCLUSION The GPC-GC/FID and GPC-GC/MS method established in the study have good specificity, high precision, accuracy and robustness, which can be used for the separation and determination of free lanolin alcohol and six organophosphorus and pyrethroid residual pesticides in lanolin. At the same time, this study can provide reference for the quality control of lanolin.


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