Aim To investigate the regulatory mecha-nisms of donepezil on the expression and enzymatic ac-tivity of cytochrome P450 3A4(CYP3A4),elucidate the synergistic impact of hypoxia on CYP3A4 function,and reveal its potential association with drug-induced cardiotoxicity,particularly QT interval prolongation.Methods Western blot,co-immunoprecipitation,and gene knockdown techniques were employed to evaluate the effects of donepezil and hypoxia on CYP3A4 pro-tein expression.CYP3A4 enzymatic activity was as-sessed using an in vitro incubation system with rat liver microsomes combined with high-performance liquid chromatography(HPLC),and the half-maximal inhib-itory concentration(IC50)was determined.Results Donepezil(10 μmol·L-1)and hypoxia reduced CYP3A4 protein expression to 31.75％and 45.90％of the control levels,respectively.Both interventions activated the gp78-mediated ubiquitin-proteasome path-way,significantly increasing CYP3A4 ubiquitination levels by 2.1-fold compared to the control group,thereby promoting proteasomal degradation.Donepezil inhibited CYP3A4 enzyme activity with an IC50 of 83.4μmol·L-1,and hypoxia synergistically enhanced this inhibitory effect,reducing the IC50 to 20.79 μmol·L-1.Conclusion Donepezil downregulates CYP3A4 function through dual mechanisms involving ubiquitin-mediated proteasomal degradation and direct enzymatic inhibition.Hypoxia potentiates this effect,leading to impaired metabolism of CYP3A4 substrate drugs,ele-vated plasma drug concentrations(1.6-2.3-fold in-crease compared to normal metabolic conditions),and an increased risk of QT interval prolongation and other forms of cardiotoxicity.

Objective:To evaluate the efficacy and safety of spesolimab in the treatment of generalized pustular psoriasis (GPP) .Methods:Six patients with GPP were retrospectively collected from Nanjing Drum Tower Hospital from February 2024 to April 2024, including 2 males and 4 females, aged 16 - 50 years. Four patients had a history of plaque psoriasis. Before treatment, the GPP Area and Severity Index (GPPASI) scores of the patients ranged from 24.6 to 60.8 points, and their GPP Physician Global Assessment (GPPGA) scores ranged from 2 to 4 points. All the 6 patients were intravenously injected with a single dose of 900 mg of spesolimab. Their GPPASI and GPPGA scores were recorded at weeks 1, 4, and 12, and adverse reactions were monitored during the treatment and follow-up.Results:Twenty-four hours after treatment, pustules were markedly reduced in the 6 patients with GPP; at week 1, pustules completely subsided, but erythema persisted; at week 4, both GPPASI and GPPGA scores decreased to 0 points; at week 12, no recurrence was observed. The patients with comorbid hypertension (4 cases), diabetes (1 case), or kidney disease (2 cases) maintained stable pressure or blood glucose levels, or showed improved renal function during the treatment and follow-up. Two patients experienced a transient fever after the treatment with spesolimab, resolving spontaneously after 2 days; two other patients exhibited exacerbation of plaque psoriasis. During the 12-week follow-up, none of the 6 patients experienced serious adverse reactions such as infections.Conclusion:In this study, spesolimab demonstrated rapid-onset efficacy and favorable safety in the treatment of GPP; conventional-dose spesolimab seemed safe for GPP patients with kidney disease.

Objective:To establish a quantitative analysis of multi-components by single marker(QAMS)for the determination of 6 alkaloid components,which is benzoylmesaconine,benzoyl-hypaconine,benzoylaconine,mesaconitine,hypaconitine,and aconitine in Zhachong Shisanwei Pills,and prove the scientificity and feasibility of the method in the quality analysis.Methods:The chromatographic separation was performed on an Agilent Eclipse Plus C18(250 mm×4.6 mm,5 μm)with gradient elution using 0.1 mol·L-1 ammonium acetate(0.5 mL of gla-cial acetic acid per 1 000 mL)(A)-acetonitrile:tetrahydrofuran(25∶15)(B),as the mobile phase(0-50 min,18%B-28%B),the detection wavelength was switched from 235 nm,the column temperature was kept at 40℃and the flow rate was 1.0 mL·min-1.The relative correction factors(fs/i)were established with the other 5 compo-nents to be measured using benzoylaconine as the internal reference,which were used to calculate the mass fraction of each component.At the same time,the mass fractions of the 6 effective constituents in Zhachong Shisanwei Pills were calculated by the external standard method(ESM).By comparing the content results of ESMand QAMS,the accura-cy of QAMS method were evaluated.Results:The relative correction factors(fs/i)of benzoylmesaconine,benzoylhyp-aconine,mesaconitine,hypaconitine,and aconitine in Mongolian medicine Zhachong Shisanwei Pills were reproduci-ble with good reproducibility,which were 0.680 4,0.450 6,0.850 8,0.676 1 and 0.757 0,the result obtained by QAMS approximated those obtained by external standard method(ESM).Conclusion:The method is simple,stable and reproducible,and can be used for the quality control of 6 alkaloid components in Zhachong Shisanwei Pills.

This study aims to comprehensively analyze the material basis of toad visceral oil(hereafter referred to as toad oil), and explore the pharmacological effect of toad oil on atopic dermatitis(AD). Ultra-high performance liquid chromatography-linear ion trap/orbitrap high-resolution mass spectrometry(UHPLC-LTQ-Orbitrap-MS) and gas chromatography-mass spectrometry(GC-MS) were employed to comprehensively identify the chemical components in toad oil. The animal model of AD was prepared by the hapten stimulation method. The modeled animals were respectively administrated with positive drug(0.1% hydrocortisone butyrate cream) and low-and high-doses(1%, 10%) of toad oil by gavage. The effect of toad oil on AD was evaluated with the AD score, ear swelling rate, spleen index, and pathological section results as indicators. A total of 99 components were identified by UHPLC-LTQ-Orbitrap-MS, including 14 bufadienolides, 7 fatty acids, 6 alkaloids, 10 ketones, 18 amides, and other compounds. After methylation of toad oil samples, a total of 20 compounds were identified by GC-MS. Compared with the model group, the low-and high-dose toad oil groups showed declined AD score, ear swelling rate, and spleen index, alleviated skin lesions, and reduced infiltrating mast cells. This study comprehensively analyzes the chemical composition and clarifies the material basis of toad oil. Meanwhile, this study proves that toad oil has a good therapeutic effect on AD and is a reserve resource of traditional Chinese medicine for external use in the treatment of AD.
Animals ; Dermatitis, Atopic/immunology* ; Disease Models, Animal ; Mice ; Male ; Gas Chromatography-Mass Spectrometry ; Humans ; Bufonidae ; Oils/administration & dosage* ; Chromatography, High Pressure Liquid ; Female ; Mice, Inbred BALB C

Trigeminal neuralgia is characterized by intense pain in the sensory distribution area of the trigeminal nerve. It can be triggered by non-noxious stimuli such as brushing teeth and washing face. At present, the treatment of trigeminal neuralgia mainly includes oral drugs and surgical treatments. A 92-year-old patient with trigeminal neuralgia was reported. The pain could not be alleviated because the patient was unable to tolerate the side effects of drugs and surgical treatment. Taking into account the onset time and the duration of the curative effect, botulinum toxin type A was combined with cobrotoxin for the treatment of the patient. As a result, the pain symptoms were rapidly alleviated and remained in a relieved state for 8 months. The clinical characteristics of this patient were summarized in this article, and the possible synergistic mechanisms of action of the 2 drugs were discussed. The ultimate objective is to furnish a broader spectrum of alternatives and references for clinical practice.

OBJECTIVES: To evaluate the application value of genetic newborn screening (gNBS) in the Yunnan region. METHODS: A prospective study was conducted with a random selection of 3 001 newborns born in the Yunnan region from February to December 2021. Traditional newborn screening (tNBS) was used to test biochemical indicators, and targeted next-generation sequencing was employed to screen 159 genes related to 156 diseases. Positive-screened newborns underwent validation and confirmation tests, and confirmed cases received standardized treatment and long-term follow-up. RESULTS: Among the 3 001 newborns, 166 (5.53%) were initially positive for genetic screening, and 1 435 (47.82%) were genetic carriers. The top ten genes with the highest variation frequency were GJB2 (21.29%), DUOX2 (7.27%), HBA (6.14%), GALC (3.63%), SLC12A3 (3.33%), HBB (3.03%), G6PD (2.94%), SLC25A13 (2.90%), PAH (2.73%), and UNC13D (2.68%). Among the initially positive newborns from tNBS and gNBS, 33 (1.10%) and 47 (1.57%) cases were confirmed, respectively. A total of 48 (1.60%) cases were confirmed using gNBS+tNBS. The receiver operating characteristic curve analysis demonstrated that the areas under the curve for tNBS, gNBS, and gNBS+tNBS in diagnosing diseases were 0.866, 0.982, and 0.968, respectively (P<0.05). DeLong's test showed that the area under the curve for gNBS and gNBS+tNBS was higher than that for tNBS (P<0.05). CONCLUSIONS gNBS can expand the range of disease detection, and its combined use with tNBS can significantly shorten diagnosis time, enabling early intervention and treatment.
Humans ; Infant, Newborn ; Neonatal Screening ; Genetic Testing ; Female ; Male ; Follow-Up Studies ; Prospective Studies ; China

OBJECTIVE: To evaluate the efficacy and safety of Shexiang Tongxin Dropping Pill (STDP) in treating stable angina patients with phlegm-heat and blood-stasis syndrome by exercise duration and metabolic equivalents. METHODS: This multicenter, randomized, double-blind, placebo-controlled clinical trial enrolled stable angina patients with phlegm-heat and blood-stasis syndrome from 22 hospitals. They were randomized 1:1 to STDP (35 mg/pill, 6 pills per day) or placebo for 56 days. The primary outcome was the exercise duration and metabolic equivalents (METs) assessed by the standard Bruce exercise treadmill test after 56 days of treatment. The secondary outcomes included the total angina symptom score, Chinese medicine (CM) symptom scores, Seattle Angina Questionnaire (SAQ) scores, changes in ST-T on electrocardiogram and adverse events (AEs). RESULTS: This trial enrolled 309 patients, including 155 and 154 in the STDP and placebo groups, respectively. STDP significantly prolonged exercise duration with an increase of 51.0 s, compared to a decrease of 12.0 s with placebo (change rate: -11.1% vs. 3.2%, P<0.01). The increase in METs was significantly greater in the STDP group than in the placebo group (change: -0.4 vs. 0.0, change rate: -5.0% vs. 0.0%, P<0.01). The improvement of total angina symptom scores (25.0% vs. 0.0%), CM symptom scores (38.7% vs. 11.8%), reduction of nitroglycerin consumption (100.0% vs. 11.3%), and all domains of SAQ, were significantly greater with STDP than placebo (all P<0.01). The changes in Q-T intervals at 28 and 56 days from baseline were similar between the two groups (both P>0.05). Twenty-five participants (16.3%) with STDP and 16 (10.5%) with placebo experienced AEs (P=0.131), with no serious AEs observed. CONCLUSION STDP could improve exercise tolerance in patients with stable angina and phlegm-heat and blood stasis syndrome, with a favorable safety profile. (Registration No. ChiCTR-IPR-15006020).
Humans ; Double-Blind Method ; Drugs, Chinese Herbal/adverse effects* ; Male ; Female ; Middle Aged ; Angina, Stable/physiopathology* ; Aged ; Syndrome ; Treatment Outcome ; Placebos ; Tablets

Objective:To evaluate the efficacy and safety of spesolimab in the treatment of generalized pustular psoriasis (GPP) .Methods:Six patients with GPP were retrospectively collected from Nanjing Drum Tower Hospital from February 2024 to April 2024, including 2 males and 4 females, aged 16 - 50 years. Four patients had a history of plaque psoriasis. Before treatment, the GPP Area and Severity Index (GPPASI) scores of the patients ranged from 24.6 to 60.8 points, and their GPP Physician Global Assessment (GPPGA) scores ranged from 2 to 4 points. All the 6 patients were intravenously injected with a single dose of 900 mg of spesolimab. Their GPPASI and GPPGA scores were recorded at weeks 1, 4, and 12, and adverse reactions were monitored during the treatment and follow-up.Results:Twenty-four hours after treatment, pustules were markedly reduced in the 6 patients with GPP; at week 1, pustules completely subsided, but erythema persisted; at week 4, both GPPASI and GPPGA scores decreased to 0 points; at week 12, no recurrence was observed. The patients with comorbid hypertension (4 cases), diabetes (1 case), or kidney disease (2 cases) maintained stable pressure or blood glucose levels, or showed improved renal function during the treatment and follow-up. Two patients experienced a transient fever after the treatment with spesolimab, resolving spontaneously after 2 days; two other patients exhibited exacerbation of plaque psoriasis. During the 12-week follow-up, none of the 6 patients experienced serious adverse reactions such as infections.Conclusion:In this study, spesolimab demonstrated rapid-onset efficacy and favorable safety in the treatment of GPP; conventional-dose spesolimab seemed safe for GPP patients with kidney disease.

Objective To explore the mechanism by which the sanguis draconis flavones(SDF)regulates the reactive oxygen species(ROS)/thioredoxin-interacting protein(TXNIP)pathway to mediate cell pyroptosis and improve cerebral ischemia-reperfusion injury(CIRI)in rats.Methods The experimental rats were randomly divided into the control group(Ctrl),the CIRI group,the low-dose SDF group(SDF-L),the high-dose SDF group(SDF-H),and the SDF-H+ROS/TXNIP pathway activator,trimethylamine oxide(TMAO)group(SDF-H+TMAO).Among them,except for the control group,the remaining rats all needed to establish the CIRI rat model by the modified suture method.Zea Longa scoring was performed on rats from each group.ELISA was used to detect the levels of serum inflammatory factors interleukin(IL)-1β,IL-18 and oxidative stress-related factors superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px).Flow cytometry was used to measure the ROS levels.Cerebral edema was detected.Cerebral infarction was detected by 2,3,5-triphenyl tetrazolium chloride(TTC)staining.HE staining was used to detect the pathological changes of brain tissue.Immunohistochemistry was used to detect the expression of pyrolytic effector protein dermolin D(GSDMD).Western blotting was used to detect the expression of proteins related to the ROS/TXNIP pathway.Results Compared with the control group,a large area of cerebral infarctions were observed in the brain tissue of the CIRI group,accompanied by mild hemorrhage and obvious infiltration of inflammatory cells.Neuronal cells underwent degeneration and necrosis,with sparse and disordered arrangement.The phenomena of nuclear condensation and nucleolus lysis were obvious.The Zea Longa score,cerebral infarction volume,brain tissue water content,levels of IL-1β,IL-18,ROS,MDA,and the expressions of GSDMD,TXNIP,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-related punctate protein(ASC),and Caspase-1 increased,while the activities of SOD and GSH-Px decreased(P＜0.05).Compared with the CIRI group,the pathological damage of brain tissues in the SDF-L group and the SDF-H group was significantly improved.The Zea Longa score,cerebral infarction volume,brain tissue water content,levels of IL-1β,IL-18,ROS,MDA,and the expressions of GSDMD,TXNIP,NLRP3,ASC,and Caspase-1 decreased.The activities of SOD and GSH-Px increased(P＜0.05);TMAO treatment partially reversed the improvement effect of SDF on CIRI in rats.Conclusion SDF ameliorates cerebral CIRI in rats by inhibiting ROS/TXNIP pathway-mediated pyroptosis.

Objective:To establish a quantitative analysis of multi-components by single marker(QAMS)for the determination of 6 alkaloid components,which is benzoylmesaconine,benzoyl-hypaconine,benzoylaconine,mesaconitine,hypaconitine,and aconitine in Zhachong Shisanwei Pills,and prove the scientificity and feasibility of the method in the quality analysis.Methods:The chromatographic separation was performed on an Agilent Eclipse Plus C18(250 mm×4.6 mm,5 μm)with gradient elution using 0.1 mol·L-1 ammonium acetate(0.5 mL of gla-cial acetic acid per 1 000 mL)(A)-acetonitrile:tetrahydrofuran(25∶15)(B),as the mobile phase(0-50 min,18%B-28%B),the detection wavelength was switched from 235 nm,the column temperature was kept at 40℃and the flow rate was 1.0 mL·min-1.The relative correction factors(fs/i)were established with the other 5 compo-nents to be measured using benzoylaconine as the internal reference,which were used to calculate the mass fraction of each component.At the same time,the mass fractions of the 6 effective constituents in Zhachong Shisanwei Pills were calculated by the external standard method(ESM).By comparing the content results of ESMand QAMS,the accura-cy of QAMS method were evaluated.Results:The relative correction factors(fs/i)of benzoylmesaconine,benzoylhyp-aconine,mesaconitine,hypaconitine,and aconitine in Mongolian medicine Zhachong Shisanwei Pills were reproduci-ble with good reproducibility,which were 0.680 4,0.450 6,0.850 8,0.676 1 and 0.757 0,the result obtained by QAMS approximated those obtained by external standard method(ESM).Conclusion:The method is simple,stable and reproducible,and can be used for the quality control of 6 alkaloid components in Zhachong Shisanwei Pills.