1.The feasibility of using high-definition thoracoscopy to identify sympathetic ganglia during thoracic sympathicotomy for primary palmar hyperhidrosis
Gang XU ; Chaoyue HU ; Cong CHEN ; Yuancai LIN ; Daolong ZHU ; Han LIU ; Dong WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(04):578-583
Objective To explore the feasibility of using high-definition thoracoscopy to identify sympathetic ganglia during thoracic sympathicotomy for primary palmar hyperhidrosis. Methods The clinical data of patients with primary palmar hyperhidrosis who underwent high-definition thoracoscopic sympathicotomy in Taikang Xianlin Drum Tower Hospital from June to July 2023 were retrospectively analyzed. Intraoperative visualization rates and anatomical variations of sympathetic ganglia were recorded, and the consistency between white-light thoracoscopy and near-infrared fluorescence imaging was compared. Additionally, surgical videos from previous fluorescence-guided procedures were reviewed. Results Finally 100 patients were collected, including 54 females and 46 males, with an average age of (21.92±6.56) years. All patients underwent endoscopic thoracic sympathicotomy at R3 level. The overall intraoperative ganglion visualization rate was 92.5% (740/800), with G2-G5 rates of 95.5% (191/200), 94.0% (188/200), 94.0% (188/200), and 86.5% (173/200), respectively. Ganglion variations occurred in 32.0% (237/740), predominantly at G3 (29.8%) and G4 (42.6%). In 5 indocyanine green-enhanced patients, the concordance rate between white-light and near-infrared fluorescence imaging was 100.0% (38/38). Video analysis of 14 near-infrared fluorescence-guided surgeries demonstrated a 99.1% (107/108) consistency rate. Postoperative palmar hyperhidrosis improvement reached 100.0% (100/100) with no Horner’s syndrome. Conclusion With the wide clinical application of high-definition thoracoscopy, accurate thoracic sympathicotomy has the feasibility of clinical application.
2.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.
3.Compact Fundus Imaging System Using Shack-Hartmann Wavefront Sensing for High-speed Auto-focus
Zhe-Kai LIN ; Long CHEN ; Geng-Yong ZHENG ; Jin-Tian HUANG ; Jia-Xin DONG ; Shang-Pan YANG ; Wen-Zheng DING ; Ding-An HAN ; Xue-Hua WANG ; Ya-Guang ZENG
Progress in Biochemistry and Biophysics 2026;53(4):1076-1086
ObjectiveThe widespread adoption of portable fundus cameras for primary care and community screening is hindered by limitations in current autofocus(AF) technologies. Image-based methods relying on sharpness evaluation require iterative searches, resulting in slow convergence, while projection-based techniques are susceptible to optical artifacts and calibration errors. To address these challenges, this study introduces a novel AF system based on direct wavefront sensing, designed to deliver simultaneous high speed, high precision, and operational robustness within the compact form factor essential for portable ophthalmic devices. MethodsOur approach fundamentally reimagines the AF process by directly measuring the ocular wavefront aberration. We developed a custom portable fundus camera integrating a miniaturized Shack-Hartmann wavefront sensor (SHWS) into the optical path. An 850 nm laser diode projects a point source onto the retina via oblique illumination to minimize corneal reflections. Light scattered from this spot carries the eye’s refractive error through the imaging optics and is directed to the SHWS, positioned at a plane optically conjugate to the primary color CMOS imaging sensor. A microlens array within the SHWS samples the incident wavefront, generating a pattern of focal spots on a CCD. Real-time centroid analysis of these spots provides a map of local wavefront slopes. These measurements are processed through a singular value decomposition (SVD) algorithm to fit a Zernike polynomial basis set, enabling real-time reconstruction of the wavefront phase. The defocus component (S) is extracted from the second-order Zernike coefficients, providing a direct, quantitative measure of the refractive error in diopters. This value serves as a precise error signal in a closed-loop control system, which commands a voice-coil actuated focusing lens to its null position in a single, deterministic step, eliminating the need for iterative search algorithms. ResultsComprehensive evaluation demonstrated the system’s high performance. Testing on a calibrated model eye (OEMI-7) established a highly linear relationship between the computed defocus S and the focusing lens position across a ±20 Diopter (D) compensation range, achievable within a 5 mm mechanical travel. The system achieved a focusing precision of 0.08 D, corresponding to an 18-fold improvement over a conventional projection spot-size method tested under identical conditions. The total focus acquisition time, encompassing wavefront measurement, computation, and lens actuation, averaged under 0.5 s. Clinical validation with 25 human volunteers (50 eyes, refractive range -15 D to +10 D) confirmed practical efficacy. The wavefront-sensing AF succeeded in 92% of attempts with a mean time of 0.5 s, substantially outperforming a projection-based benchmark which achieved only a 32% success rate with an average time of 4.25 s. The system provided instantaneous directional guidance and maintained stability during minor ocular movements. Objective assessment of image quality, via amplitude contrast of retinal vasculature, showed consistent and significant enhancement following AF correction across the entire tested diopter range. ConclusionThis work successfully implements and validates a direct wavefront-sensing autofocus paradigm for portable fundus cameras. By directly quantifying and compensating for the optical defocus aberration, this method bypasses the fundamental limitations of image-processing and projection-based techniques, enabling rapid, precise, and deterministic diopter compensation. The developed system delivers an exceptional combination of a wide operational range (±20 D), high accuracy (0.08 D), fast convergence (0.5 s), and a compact physical footprint. This technology provides a practical and high-performance focusing solution capable of enhancing the reliability, throughput, and diagnostic utility of portable retinal imaging in large-scale screening applications. Future efforts will be directed towards system cost optimization and performance adaptation for diverse ocular conditions.
4.Confirmatory analysis of HBsAg reactive samples from voluntary blood donors
Qiaolin ZHANG ; Fang WANG ; Dong LIU ; Fengjiao HAN ; Liu LI ; Xiaochuan ZHENG ; Xuelian DENG ; Dongyan YANG
Chinese Journal of Blood Transfusion 2026;39(4):452-457
Objective: To systematically analyze the confirmatory positivity of different combinations of HBsAg screening results in blood testing, providing data to support the optimization of blood donor eligibility management. Methods: A retrospective analysis was conducted on blood screening data from 174 266 voluntary blood donor samples at the Chongqing Blood Center between October 2021 and September 2022. Samples with inconsistent results between the two HBsAg enzymelinked immunosorbent assays (ELISA) and individual donor nucleic acid testing (NAT) were confirmed using an electrochemiluminescence immunoassay (ECLIA) and a neutralization test. The detection efficacy of four different HBsAg ELISA reagents was compared using the HBsAg-confirmed positive samples. Results: A total of 767(0.44%) HBV-reactive (HB-sAg and/or HBV DNA reactive) samples were detected. Among them, 344 samples with discordant serological and NAT results were collected, of which 64(18.6%) were confirmed positive by neutralization test. Additionally, 5 samples that were neutralization-negative but double-reactive for HBsAg and HBV DNA were confirmed as positive according to FDA guidance, resulting in a total of 69(20.1%) confirmed HBsAg-positive samples. There were significant differences in the neutralization test confirmation rates among different screening result categories (P<0.05): The group with dual HBsAg reagent reactivity (double reactive) & NAT-negative had the highest confirmation rate (96.9%, 31/32); the group reactive to only reagent 2 (single reactive) had a rate of 25.7% (29/113); while the confirmation rates for samples reactive to only reagent 1 and samples with isolated HBV DNA positivity were extremely low [0(0/34) and 2.4%(4/165), respectively]. The four commercial reagents showed significant differences in their ability to detect confirmed positive samples that were initially single reactive (P<0.05). Conclusion: Given the performance variations among HBsAg screening reagents, thorough performance verification is essential before implementation. When NAT is negative, dual HBsAg reactivity in screening can serve as a basis for confirming infection and directly deferring blood donors. However, confirming infection in donors with single HBsAg reactivity is more challenging, necessitating supplementary tests to rule out infection risk.
5.The magnetic compression technique in the treatment of severe mid-ureteral stricture
Fanchao WEI ; Zhaoxiang WANG ; Shichao HAN ; Zhiwen DONG ; Yuxin FU ; Ruochen QI ; Guohui WANG ; Xiaoyan ZHANG ; Tong XU ; Jingliang ZHANG ; Weijun QIN ; Lijun YANG ; Shuaijun MA
Journal of Modern Urology 2026;31(2):182-186
Objective To explore the feasibility of magnetic compression technique (MCR) in the treatment of severe single mid-ureteral stricture in a rabbit model. Methods After 10 female New Zealand rabbits were selected, the distal end of the right ureter was ligated with silk thread. Two weeks after the operation, when the right ureter was dilated, the mid-ureter was ligated again with silk thread to create a severe single mid-ureteral stricture animal model. Longitudinal incisions were made at the upper and lower ends of the ligation site, and cylindrical NdFeB magnets with central holes (sub-magnets and main magnets) were placed to anastomose and recanalize the mid-ureteral stricture. The magnets were adjusted to the appropriate position to automatically align and attract each other. The longitudinal incision sites of the ureter were sutured with 6-0 Prolene thread, and the abdominal cavity was closed layer by layer. The operation time was recorded. X-ray films were taken every two days to confirm the position of the magnets and record the time of magnet detachment. Three weeks after the operation, the anastomotic specimens were obtained, and the formation of the anastomosis was evaluated with gross observation and histological staining. Results Two weeks after ligation, all experimental rabbits showed obvious dilation of the right ureter. The mid-ureteral ligation and placement of the male and female magnets were successfully completed in all 10 experimental rabbits, with an operation time of (24.00±5.15)minutes. The male and female magnets attracted each other well. X-ray examination showed that the magnets detached and entered the distal ureter (14.0±1.9) days after magnetic compression. The recanalization of the severe single mid-ureteral stricture was successfully achieved. The gross specimens and histological examination of the anastomosis showed that the anastomosis remained patent and the mucosal healing was good. All experimental rabbits survived well after the operation without serious complications. Conclusion MCT provides a new idea and novel method for the treatment of mid-ureteral stricture. This study confirmed the feasibility of this technique in treating severe single mid-ureteral stricture in a rabbit model. The operation is relatively simple and has potential for clinical application.
6.Comparison of the efficacy and prognosis of dapagliflozin versus empagliflozin in elderly patients with heart failure complicated by cardiac pacemaker implantation
Bo WANG ; Ou ZHANG ; Jie LI ; Dong HAN
China Pharmacy 2026;37(16):2166-2170
OBJECTIVE To compare the efficacy and prognosis between dapagliflozin and empagliflozin in elderly patients with heart failure (HF) complicated by cardiac pacemaker implantation.METHODS The clinical data of 237 elderly patients with HF complicated by cardiac pacemaker implantation, who were admitted to the Department of Cardiovascular Medicine at the First Affiliated Hospital of Nanyang Medical College from July 1, 2023, to January 1, 2025, were retrospectively collected. According to medication use, the patients were divided into the control group (conventional therapy, n =90), dapagliflozin group (conventional therapy plus dapagliflozin, n =77), and empagliflozin group (conventional therapy plus empagliflozin, n =70). Cardiac function and HF-related indicators were evaluated before treatment and after 6 months of treatment among the 3 groups. The incidence of major adverse cardiovascular events (MACE), HF rehospitalization rate, and the occurance of adverse reaction were compared among the 3 groups.RESULTS After 6 months of treatment, compared with the same group before treatment, left ventricular ejection fraction was significantly increased, 6-minute walk distance was significantly prolonged, left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly shortened, and N-terminal pro-B-type natriuretic peptide and high-sensitivity C-reactive protein levels were significantly decreased in all 3 groups ( P <0.05). The improvements in most of the above indicators were greater in the dapagliflozin and empagliflozin groups than in the control group ( P <0.05), and were all greater in the empagliflozin group than in the dapagliflozin group ( P <0.05). The HF rehospitalization rate was significantly lower in both dapagliflozin and empagliflozin groups than in the control group ( P <0.05), and was significantly lower in the empagliflozin group than in the dapagliflozin group ( P <0.05). There was no statistically significant difference in the incidence of MACE among the 3 groups ( P >0.05), although the incidence was numerically lower in the dapagliflozin and empagliflozin groups than in the control group. There was also no statistically significant difference in the incidence of adverse reactions among the 3 groups ( P >0.05).CONCLUSIONS Both dapagliflozin and empagliflozin can improve cardiac function and HF-related indicators and reduce the risk of HF rehospitalization in elderly patients with HF complicated by cardiac pacemaker implantation on the basis of conventional therapy, without increasing the risk of adverse reactions. Compared with dapagliflozin, empagliflozin shows more pronounced effects.
7.Analysis of knowledge, attitude, and practice levels regarding health protection against air pollution and related factors among Chinese primary school students
Chinese Journal of School Health 2026;47(7):958-961
Objective:
To assess the current status of knowledge, attitude, and practice (KAP) regarding health protection against air pollution among primary school students in China, and to analyze the impacts of fine particulate matter (PM 2.5 ) exposure, family, and individual factors on KAP levels, so as to provide empirical basis for implementing targeted interventions.
Methods:
Using stratified cluster random sampling method, a questionnaire survey was conducted among 19 297 primary school students from third to fifth grade across 30 provinces (autonomous regions and municipalities directly under the central government) in China in 2024. The questionnaire was developed based on the Technical Guidelines for the Assessment of Civic Environmental and Health Literacy (Trial), and encompasses three dimensions:knowledge, attitude, and practice. Spearman correlation analysis, Chi-square test, multivariate Logistic regression, and Bootstrap mediation effect test was applied for data analysis.
Results:
The overall mean KAP score for health protection against air pollution among primary school students was (79.73±13.53), with an adequacy rate of 80.47% ( n =15 528). The dimension scores for knowledge, attitude, and practice were (41.34±8.01, 16.43± 2.61 ), and (21.97±5.71), respectively. Knowledge was positively correlated with attitude and practice, and attitude was positively correlated with practice ( r =0.51, 0.35, 0.32, all P <0.01). Multivariate Logistic regression showed that the northeast region ( OR =2.36, 95% CI =2.02-2.75), having parents with a bachelor s degree or higher ( OR =4.76, 95% CI =2.71-8.37), living with both parents ( OR =1.33, 95% CI =1.15-1.55), being in the fifth grade ( OR =1.22, 95% CI =1.11-1.33), outdoor PM 2.5 level exceeding standard limits at the survey site ( OR =1.25, 95% CI =1.16-1.34), history of allergy ( OR =1.47, 95% CI =1.31-1.64), and wearing masks during heavily polluted weather ( OR =1.19, 95% CI =1.07-1.31) were promoting factors for KAP adequacy rate (all P <0.05). Conversely, obesity was a detrimental factor ( OR =0.78, 95% CI =0.69-0.88, all P <0.05).
Conclusions
Primary school students in China exhibit a favorable KAP level regarding health protection against air pollution, though regional, familial, and individual disparities persist. Exposure to PM 2.5 levels exceeding standard limits exerts a direct positive effect on enhancing health protection literacy. Future efforts should focus on the western region, families with lower educational attainment children to implement tailored, stratified interventions.
8.Phacoemulsification on intraocular anatomical structures in cataract patients with high myopia
Kaili TANG ; Jing WANG ; Zhenbo ZHAO ; Dong HAN ; Yuxi DING ; Jikun YANG ; Jinsong ZHANG ; Liwei MA
International Eye Science 2026;26(10):1753-1758
Phacoemulsification represents the mainstream, efficient, and safe surgical modality for cataract treatment, and continuous technological advances have further improved the safety profile of this procedure in ordinary patients. However, patients with high myopia feature inherent anatomical abnormalities of the eyeball, leading to a fundamental incompatibility with conventional phacoemulsification protocols. Such patients face a markedly elevated risk of both anterior and posterior segment perioperative complications, posing a persistent clinical challenge in cataract surgery. Focusing on the unique ocular anatomical characteristics associated with high myopia, this review systematically summarizes the distinct traumatic effects and pathological impacts of phacoemulsification on the anterior and posterior ocular segments of affected patients. Moving beyond generalized discussions of routine cataract management, it consolidates clinical evidence specific to this special-population patients, aiming to provide refined references for perioperative risk prevention and control in cataract patients complicated with high myopia.
9.Development of the "Clinical Ethics" Course for the Eight-Year Clinical Medicine Program: Explorations and Practices by an Interdisciplinary Teaching Team
Jiangli HAN ; Yali CONG ; Qiong BAI ; Shu LI ; Guangkuan XIE ; Tongyan HAN ; Chuan MA ; Ying YAO ; Fei DONG ; Huilin LIU ; Rui WU ; Qinggang GE ; Liang WANG ; Yuxi XIA ; Shixian GU ; Xuan HE ; Ning SHEN
Medical Journal of Peking Union Medical College Hospital 2026;17(5):1223-1229
With the rapid advancement of medical technology and the increasing complexity of clinical environments, competence in clinical ethical decision-making has become an indispensable core competency for exemplary physicians. However, traditional medical ethics education has long been confronted with the persistent challenges of theory-practice dissociation and the disconnect between pedagogical settings and clinical realities. Thus, how to construct a "Clinical Ethics" curriculum that aligns with the objectives of training high-caliber medical professionals has emerged as a critical and challenging issue in current medical education reform. To effectively address this challenge, the Office of Education at Peking University Third Hospital, since 2021, has assembled a multidisciplinary "Clinical Ethics" teaching and management team comprising faculty from Peking University School of Humanities and key clinical specialists from the hospital. In 2022, the "Clinical Ethics" elective course was formally launched for fourth-year students in the eight-year MD program who had completed coursework in "Medical Ethics" and organ-system-based curricula, with the aim of exploring and operationalizing a pre-clinical ethics education model tailored for senior medical students. The course employs a modular instructional design encompassing "theoretical review-case-based deliberation-clinical workshop", which effectively facilitates the deep integration of medical humanities with clinical practice.Following four years of iterative refinement through the "plan-act-observe-reflect" cycle, the course has demonstrated positive outcomes in enhancing students' ethical awareness and decision-making capabilities, and has preliminarily established a sustainable collaborative mechanism among the multidisciplinary faculty team. This article systematically delineates the course development background, pedagogical model and methodological innovations, and the multidisciplinary collaborative framework, alongside reflections and summaries based on implementation outcomes, with the intent of providing references for the future development of "Clinical Ethics" curricula.
10.Gantou Fumu Decoction Inhibits Hepatic Stellate Cell Activation by Regulating Nrf2/ARE Signaling Pathway
Wangsheng HE ; Yanping ZHU ; Shizhen LONG ; Mengmeng LI ; Wenming YANG ; Han WANG ; Meixia WANG ; Ting DONG ; Shupei JIA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(23):105-115
ObjectiveTo explore the mechanism through which Gantou Fumu decoction (GDFMT)-containing serum inhibits the activation of hepatic stellate cells. MethodsSerum pharmacology techniques were employed to prepare GDFMT-containing serum samples. Rat hepatic stellate cells (HSCT6) were randomly allocated into three groups: Control, penicillamine (90 mg·kg-1), and GDFMT (6.8 g·kg-1). Cell viability was assessed by the cell counting kti-8 (CCK-8) assay, intracellular ROS levels were determined by flow cytometry using the DCFH‑DA fluorescent probe and while extracellular collagen Ⅰ (ColⅠ) levels were quantified via enzyme linked immunosorbent assay (ELISA). Intracellular α-smooth muscle actin (α-SMA) expression was visualized through immunofluorescence staining. Real-time polymerase chain reaction(Real-time PCR) and Western blot were performed to evaluate alterations in the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2), glutamate cysteine ligase, catalytic (GCLC), glutamate-cysteine ligase, modifier subunit (GCLM), Kelch-like ECH-associated protein 1 (Keap1), NAD(P)H:Quinone oxidoreductase 1 (NQO1), and heme oxygenase-1 (HO-1). Genetic knockdown of Nrf2 was achieved through small interfering (siRNA) transfection to validate pathway specificity. ResultsCCK-8 data indicated that under the selected experimental conditions, culturing HSCT6 cells with 10% (volume fraction) GDFMT-containing serum for 48 h demonstrated equivalent effect in inhibiting HSCT6 cell proliferation compared with culturing HSCT6 cells with 10% (volume fraction) penicillamine-containing serum for 36 h, with no statistically significant difference between the two groups. After treatment of HSCT6 cells with siRNA-mediated Nrf2 gene transfection technique, the inhibitory effect of GDFMT on the activation process of HSCT6 cells was significantly weakened. After HSCT6 cells were cultured with GDFMT-containing serum for 48 h, the flow cytometry with a fluorescence probe showed that the content of reactive oxygen species (ROS) in the cells was significantly reduced. After the expression level of the Nrf2 gene in HSCT6 cells was knocked down by siRNA transfection, the content of ROS increased (P<0.01), indicating that GDFMT significantly reduced the ROS level in activated HSCT6 cells, and this effect strictly depended on the integrity of the Nrf2 pathway. Quantitative analysis by ELISA showed that the synthesis of ColⅠ in the HSCT6 cell culture system treated with GDFMT-containing serum for 48 h was lower than that in the control group (P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the ColⅠ concentration in the culture supernatant increased (P<0.01). The results of α-SMA detection by IF method showed that compared with the normal group, GDFMT-containing serum reduced the expression of α-SMA (P<0.01) and the protein level of α-SMA in HSCT6 cells was increased (P<0.01) after siRNA-mediated Nrf2 silencing, suggesting that Nrf2 signaling pathway may be involved in regulating the phenotypic transformation process of the cells. Real-time PCR and Western blot results showed that GDFMT-containing serum reduced the protein level of α-SMA (P<0.01) and enhanced the gene and protein levels of Nrf2 in the nucleus and GCLC, HO-1, Keap1, GCLM, and NQO1 in the cytoplasm of HSCT6 cells (P<0.05, P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the protein level of α-SMA was increased (P<0.01), and the levels of GCLC, GCLM, Keap1, NQO1, and HO-1 were decreased (P<0.05, P<0.01). These results indicated that GDFMT-containing serum successfully triggered the Nrf2 signaling pathway of HSCT6 cells. ConclusionThe anti-hepatic fibrosis effect of 10%GDFMT was equivalent to that of penicillamine. In addition, GDFMT can inhibit HSC activation and reduce oxidative stress damage by activating the Nrf2/antioxidant response element (ARE) pathway, as manifested by the up-regulated expression of Nrf2 and downstream antioxidant genes (GCLC, GCLM, HO-1, Keap1, and NQO1) and reduced secretion of ROS, α-SMA, and ColⅠ.


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