Glycyrrhizin (GC) is a major bioactive saponin in licorice plants (Glycyrrhiza spp.). A GC-specific single-chain variable fragment (GC-scFv) was constructed from anti-GC monoclonal antibody 2H2. To obtain a soluble recombinant recognition reagent for GC immunoassays, GC-scFv was expressed using a baculovirus–insect cell platform. The variable heavy- and light-chain genes were amplified from 2H2 hybridoma-derived complementary DNA (cDNA) and assembled using a (Gly4Ser)3 linker. The construct was transferred through pET-28a(+) and pCold I into pFastBac1Mel, which contains the honeybee melittin signal sequence for secretion in Sf9 insect cells. GC-scFv was recovered by nickel-chelate affinity chromatography and characterized using indirect competitive enzyme-linked immunosorbent assay (icELISA). The calibration curve was analyzed by semi-logarithmic regression using triplicate wells at each concentration. An anti-T7-tag Western blot detected GC-scFv at approximately 29 kDa in the eluate recovered from culture medium. Free GC produced concentration-dependent inhibition, with a half-maximal inhibitory concentration (IC50) of 0.46 μg/mL and a semi-logarithmic calibration interval of 0.0488–3.125 μg/mL. The apparent limit of detection and apparent limit of quantification, defined as 10
Type 2 diabetes mellitus (T2DM) is frequently complicated by neuropathy, impaired wound healing, and poor glycemic control, creating significant clinical and economic burdens. Adjunctive modalities such as acupuncture and photobiomodulation have gained interest for their potential to support glycemic control and improve diabetes-related complications. This case series examined the therapeutic potential of a synergistic technique combining Class 3B laser photobiomodulation with traditional needle acupuncture. Eleven patients with long-standing T2DM were treated using combined protocols, with outcomes assessed through fasting plasma glucose, HbA1c, neuropathic symptom relief, and wound recovery. Patients demonstrated consistent improvements in glycemic regulation, neuropathic symptoms, sleep quality, and wound healing. Fasting plasma glucose reductions ranged from 36 to 64
Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by motor impairment, degeneration of nigrostriatal dopaminergic neurons, and pathological α-synuclein accumulation. Current therapies mainly provide symptomatic relief, while disease-modifying treatments remain limited. This study investigated the neuroprotective effects of a traditional medicine formulation composed of Calculus Bovis, Fel Ursi, and Moschus (BUM) in experimental models of PD. The efficacy of BUM pharmacopuncture solution was evaluated in an MPTP-induced C57BL/6N mouse model and MPP⁺-induced Caenorhabditis elegans models. Motor function in mice was assessed using the rota-rod, pole, and grip strength tests. Nigrostriatal dopaminergic integrity was examined by tyrosine hydroxylase (TH) immunostaining in the substantia nigra pars compacta and striatum. In C. elegans, dopaminergic neurodegeneration and α-synuclein accumulation were assessed using BZ555 and NL5901 transgenic strains, respectively. BUM significantly improved motor performance in MPTP-treated mice, as evidenced by increased rota-rod latency, reduced pole descent time, and enhanced grip strength. BUM also preserved TH-positive dopaminergic neurons in the substantia nigra and dopaminergic fibers in the striatum. The magnitude of these effects was comparable to that of levodopa. In C. elegans, BUM significantly protected dopaminergic neurons against MPP⁺-induced degeneration and reduced α-synuclein-associated fluorescence. BUM alleviates motor dysfunction, mitigates nigrostriatal dopaminergic neurodegeneration, and reduces α-synuclein accumulation across both vertebrate and invertebrate models of PD. These findings suggest that the BUM multicomponent formulation exerts multitargeted protective effects against PD-related pathology, providing a strong rationale for further mechanistic and translational investigations in neurodegenerative disease research.
This letter provides a critical appraisal of the case report titled “Auricular Acupuncture Effect on Facial Contraction via Middle Ear Muscles Reflexes.” The original report attributes the observed facial contractions to middle-ear muscle reflexes triggered by auricular acupuncture. However, several aspects of the report and its supplementary videos warrant further clarification. Review of the provided materials suggests that the observed facial contractions may have been elicited by high-intensity auditory stimuli rather than by acupuncture itself. Visual analysis indicates that the contractions may originate from the masseter muscle. Due to fascial continuity between the masseter and regional mimic muscles, such contractions may mimic true facial muscle activity. Additionally, alternative diagnostic entities—including hemimasticatory spasm and hemifacial spasm—should be considered, as both can present with unilateral involuntary contractions. Differentiation between masseteric and mimic muscle involvement is essential for accurate interpretation. Clinical examination methods such as palpation, along with electrophysiological techniques including electromyography and electroneurography, may provide valuable diagnostic clarification. Furthermore, confounding clinical factors—such as prior esthetic interventions and occlusal or dental abnormalities—should be taken into account, given their potential to influence masseter muscle activity. If the observed contractions originated from the masseter muscle, the hypothesis attributing them to middle-ear muscle reflexes may require reconsideration. This commentary emphasizes the need for clearer differentiation of underlying mechanisms and encourages additional clinical and electrophysiological evaluation to enhance diagnostic accuracy.
Acute bronchitis is a common respiratory disease characterized by acute airway inflammation, requiring effective and safe anti-inflammatory therapies. ALC-2203 is a botanical drug candidate combining Coptidis rhizoma dry extract and ivy leaf (hederacoside C) ethanolic dry extract, designed to provide complementary anti-inflammatory and symptom-relieving effects. This study aimed to evaluate the preclinical anti-inflammatory efficacy of ALC-2203 in a lipopolysaccharide (LPS)-induced mouse model, develop a quality-controlled oral tablet formulation, and assess its clinical efficacy and safety in patients with acute bronchitis. Preclinically, acute bronchitis was induced in ICR mice via intranasal LPS administration. ALC-2203 (20, 40, and 80 mg/kg) orally administered to evaluate inflammatory cells in bronchoalveolar lavage fluid (BALF) inflammatory cells, cytokines, and lung histopathology. A 318-mg tablet formulation was developed and validated using an HPLC method for berberine quantification. Clinically, a randomized, double-blind, placebo- and active-controlled, multicenter Phase 2 trial enrolled 168 patients who received placebo, ALC-2203-2 (low dose), ALC-2203-1 (medium dose), and the active control, ALC-2203-AC (Synatura® syrup), for 7 days. The primary endpoint was the change from baseline in the Bronchitis Severity Score (BSS) at Day 7. In the mouse model, ALC-2203 reduced inflammatory cells, including macrophages, neutrophils, and lymphocytes, in BALF, thereby suppressing pro-inflammatory cytokines (i.e., TNF-α, IL-1β, and IL-6) and improving pulmonary histopathology. The tablet formulation demonstrated analytical performance, including linearity (R2 ≥ 0.999), accuracy (98–102
