Polydopamine-functionalized 3D-printed scaffold co-loaded with resveratrol and NGF for enhanced neuronal differentiation under inflammatory stress.

Bifunctional peptide-conjugated vertically aligned tubular nanofibrous conduits for enhanced vascularization and nerve regeneration.

p75 neurotrophin receptor exacerbates neuronal apoptosis after intracerebral hemorrhage by promoting OTUB1 ubiquitination and degradation.

Sensory innervation of developing organs is influenced by molecular cues secreted from surrounding tissues, yet the mechanisms coordinating this tissue-tissue communication are not well understood. Tooth innervation during root development provides a valuable model to investigate how local mesenchymal cues regulate axonal growth under physiological conditions, as innervation begins and progresses alongside tooth root formation. Here we identify the histone demethylase KDM6B, expressed in cranial neural crest-derived dental mesenchyme, as a critical extrinsic regulator of tooth sensory innervation. Loss of Kdm6b in dental mesenchyme severely impairs trigeminal axon entry and branching into the dental pulp, leading to tooth root development defects. Mechanistically, loss of Kdm6b reduces the expression of bone morphogenetic protein (BMP) pathway antagonist Bambi in the dental mesenchyme by modulating H3K27me3 chromatin marks, causing overactivation of BMP signaling, which then directly suppresses the expression of nerve growth factor (Ngf). Compromised NGF activity thereby diminishes mesenchymal support for sensory axon extension during tooth root development. Haploinsufficiency of Ezh2, which antagonizes Kdm6b, or Bmpr1a, a key BMP receptor, partially rescues Ngf expression, sensory innervation, and tooth root development defects in Kdm6b mutants. Together, these findings reveal that epigenetic regulation within mesenchymal cells governs sensory innervation during organogenesis, uncovering important regulatory mechanisms that may inform future strategies for restoring innervation in tissue regenerative approaches.

Tanshinone IIA promotes mitophagy by inhibiting the MARCHF5-mediated ubiquitination and degradation of FUNDC1 to alleviate spinal cord injury.

Depressive disorder (DD) is a widespread mental illness that lacks objective diagnostic biomarkers, complicating early detection and personalized treatment. This study investigated the diagnostic value of serum interferon-gamma (IFN-γ), nerve growth factor (NGF), and their ratio, alongside thyroid peroxidase antibody (TPOAb) and glial fibrillary acidic protein (GFAP), in patients with DD compared to healthy controls. A total of 238 participants (118 with DD and 120 controls) were enrolled. Depression severity was assessed using DSM-5 and HAM-D criteria. Serum biomarkers were measured using enzyme-linked immunosorbent assays (ELISA), and receiver operating characteristic (ROC) analysis was performed to assess diagnostic performance. DD patients exhibited significantly lower IFN-γ and higher NGF levels than controls (both p < 0.001), resulting in a markedly reduced IFN-γ/NGF ratio. The IFN-γ/NGF ratio achieved the highest diagnostic accuracy (AUC = 0.858, sensitivity = 82.20%, specificity = 77.50%), outperforming IFN-γ (AUC = 0.766) and NGF (AUC = 0.848) alone. TPOAb and GFAP levels did not differ significantly between groups. The IFN-γ/NGF ratio is a promising biomarker for depressive disorder, offering superior diagnostic accuracy over individual immune or neurotrophic markers. This composite index may support more objective and biologically informed diagnosis in clinical psychiatry.

To identify proteomic signatures underlying 3-year progression from impaired fasting glucose (IFG) to diabetes. We examined IFG progression in the Atherosclerosis Risk in Communities (ARIC) study from visit 2 (1990-1992) to visit 3 (1993-1995). We tested associations of 4,955 plasma proteins (SomaScan version 4.0) with ∼3-year progression from IFG (FG 100-125 mg/dL without diabetes) to diabetes (diagnosis, medication, or FG ≥126 mg/dL) using logistic regression models adjusted for demographics, cardiometabolic risk factors, and baseline glucose with Bonferroni correction (P < 10-5). We explored biological pathways enriched among the top proteins and calculated improvements in prediction (ΔAUC and net reclassification using 3-year risk thresholds of 6% and 15% in 80% training and 20% internal validation subsamples). We validated results in the Multi-Ethnic Study of Atherosclerosis (MESA) cohort. There were 3,786 ARIC participants with IFG (mean [SD] age 57 [6] years, 52% female, 28% Black individuals). The 3-year cumulative incidence of diabetes was 6%. Six proteins were associated with ∼3-year progression to diabetes, namely lower receptor-type tyrosine-protein phosphatase S (PTPRS), anthrax toxin receptor 2 (ANTXR2), adiponectin (ADIPOQ), ciliary neurotrophic factor receptor subunit α (CNTFR), transmembrane protein 132C (TMEM132C), and higher ADAMTS-like protein 2 (ADAMTSL2). Altered carbohydrate metabolism and glycolysis were key pathways. Adding the six proteins to covariates improved discrimination (optimism-corrected AUC 0.81, ΔAUC 0.03, P = 0.005) and net reclassification (training 12.2%, internal validation 12.0%) with predicted diabetes risk quintiles spanning <1% to ∼20%. Two of the six proteins were validated in MESA (P < 0.008). We identified proteins associated with 3-year IFG progression, with improvements in diabetes risk stratification.

A randomized, controlled, experimental animal study. This study investigates the influence ofHypericum perforatum (St. John's Wort) and its active compound, hypericin, influence on oxidative stress markers and neurotrophic factor expression during the acute phase of SCI in male rats. University animal research facility equipped with dedicated surgical suites for SCI induction and controlled-environment housing for rodents. SCI was induced in Wistar rats through a contusion injury model. The treatment groups received either hypericin (50/100 mg/kg, i.p.) or Hypericum perforatum extract (50/100 mg/kg, i.p.) for two weeks. Motor function was evaluated with the Basso, Beattie, and Bresnahan (BBB) test, and nociceptive sensitivity was assessed using the tail flick test. Serum levels of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), nitric oxide (NO), and reactive oxygen species (ROS) were measured. Histological analysis of myelination was conducted using Luxol fast blue staining. SCI led to a significant decline in BBB scores, an increase in oxidative stress markers (NO, ROS), and a reduction in BDNF levels. Treatment with 100 mg/kg hypericin notably improved BBB scores (p < 0.00001), reduced NO and ROS levels (p < 0.0001), and increased BDNF expression compared to untreated SCI rats. Histological examination showed less demyelination in the treated groups, with hypericin demonstrating more pronounced effects than Hypericum perforatum extract. Hypericin exhibited strong neuroprotective properties by reducing oxidative stress, enhancing neurotrophic factor expression, and preserving myelin integrity. These findings highlight its potential as a therapeutic agent for reducing secondary damage following SCI.

BackgroundTropomyosin receptor kinases (TRK) TRKA, TRKB and TRKC (encoded by NTRK1, NTRK2 and NTRK3, respectively), regulate survival, proliferation and differentiation of neurons through ligand-dependent activation of downstream signaling pathways including MAPK, PI3K and PKC. TRK-fusion positive cancers have ligand-independent activation of TRKs and have emerged as potent oncogenic drivers that can be very effectively targeted by FDA-approved TRK inhibitors. In intrahepatic cholangiocarcinoma (iCCA), however, NTRK fusions account only for a very small subset (<1%) and TRK-inhibitors have therefore been little explored in iCCA. The ligands of TRKs are neurotrophins, members of the nerve growth factor family, which include neurotrophin-3 (NTF3) and brain-derived neurotrophic factor (BDNF). NTF3 is the high-affinity ligand of TRKC but can bind to TRKA, TRKB and TRKC while BDNF is the high-affinity ligand of TRKB. As we recently demonstrated that hepatic stellate cells (HSCs), which are precursors of cancer-associated fibroblasts, release NTF3, promoting hepatocyte proliferation via paracrine TRKB signaling, we investigated if TRK signaling is activated in iCCA to assess the therapeutic potential of pharmacologic TRK inhibition.MethodsGene expression of NTF3, BDNF, NTRK1, NTRK2 and NTRK3 were assessed in publicly available iCCA datasets. Human iCCA specimens were analyzed by western blotting and immunohistochemistry for protein expression of TRK receptors and their ligands. NTF3- or BDNF-dependent cell proliferation was studied in human iCCA cell lines HuCCT1, Huh28 and liver fibroblast cell line LX2. Larotrectinib and taletrectinib, FDA-approved TRK inhibitors, were used for inhibition studies. iCCA patient-derived organoid lines were generated and used to study NTF3- or BDNF-dependent EdU uptake by immunofluorescence.ResultsAnalysis of publicly available transcriptomic CCA datasets showed significant overexpression of NTRK1, NTRK2, BDNF in CCAs compared to normal livers. Detailed transcriptomic analysis in another dataset of 122 patients showed highest enrichment of NTRK1, NTRK2, NTRK3, BDNF or NTF3 in the ‘immune classical’ and ‘inflammatory stroma’ subtype which are characterized by more abundant stroma and inflammation representing approximately 36% (44/122) of patients in this dataset. Similarly, western blot analysis of paired iCCA and adjacent non-tumorous liver tissue demonstrated increased expression of TRKB, TRKC, and NTF3 in a subset of human iCCA samples. Co-immunofluorescence analysis with reelin, a CAF marker, demonstrated increased expression of BDNF and NTF3 in CAFs. Incubation of HuCCT1, Huh28 and LX2 cells with BDNF or NTF3 resulted in a significant increase in cellular proliferation, which was inhibited by co-incubation with larotrectinib or taletrectinib. Consistently, BDNF and NTF3 induced increased EdU uptake in iCCA PDOs which was significantly reduced following co-incubation with larotrectinib.ConclusionThese findings demonstrated that TRKB, TRKC and NTF3 are overexpressed in human iCCA. Moreover, NTF3 and BDNF induced proliferation in iCCA and HSC cell lines, as well as iCCA PDOs, which was significantly decreased by co-incubation with TRK inhibitors larotrectinib or taletrectinib. Collectively, our data support TRK signaling as a growth signaling pathway in iCCA which merits further investigation of pan-TRK inhibitors as a potential therapeutic strategy for iCCA.

Inadvertent p75NTR signaling might cause inconsistencies in the neuroprotection offered by mesenchymal stem cells.

Overactive bladder (OAB) is a prevalent symptom-based syndrome lacking objective diagnostic tests. Urinary biomarkers, particularly nerve growth factor (NGF), have emerged as potential non-invasive tool for diagnosis and treatment monitoring. To systematically review and meta-analyze the association between urinary biomarker with treatment response and symptom score changes in adults with OAB. Prospective clinical studies relating urinary biomarkers to established OAB treatments were identified in eight databases up to May 2024, selected with predefined criteria, assessed with the Newcastle-Ottawa Scale (NOS), and synthesized as percentage mean changes; meta-analysis was limited to NGF. Nine studies carried out NGF data and smaller subsets reported on other biomarkers, showing post-treatment biomarker reductions in both responders and non-responders, with greater decreases and significantly larger symptom score improvements in responders. Evidence regarding the usefulness of urinary biomarkers in OAB patients is still heterogeneous and limited. Urinary NGF levels decrease during OAB treatment in accordance with symptoms questionnaires scores improvement suggesting that some urinary biomarkers may serve as a useful tool for monitoring treatment response in patients for whom symptom questionnaires cannot be administered or are considered unreliable, contributing in the differentiation of different disease phenotypes.

Schwann cell derived extracellular vesicles (SC-EVs) have emerged as a specialized extracellular vesicle subtype with intrinsic neuroregenerative, neuroimmune, and neurovascular regulatory capacities, making them particularly relevant for diabetic oral mucosal wound healing. Unlike more widely studied extracellular vesicles derived from mesenchymal stem cells, SC-EVs originate from the principal glial cells of the peripheral nervous system and therefore retain biological programs directly related to axonal repair, neurotrophic support, and nerve-associated tissue regeneration. Diabetic oral mucosal wounds are characterized by persistent inflammation, impaired angiogenesis, oxidative stress, extracellular matrix dysregulation, and neuropathy-associated defects in epithelial repair. SC-EVs contain a distinctive cargo of proteins, RNAs, and lipids, including neurotrophic factors such as nerve growth factor and brain-derived neurotrophic factor, regulatory microRNAs such as miR-21 and miR-146a, and antioxidant molecules that collectively modulate these pathological processes. This review first summarizes the pathophysiological features of diabetic oral mucosal wounds, with emphasis on neurovascular and neuroimmune disruption. It then discusses the biogenesis, molecular composition, and functional mechanisms of SC-EVs, highlighting their roles in inflammatory recalibration, angiogenesis, neuroregeneration, and redox homeostasis. A dedicated discussion of SC-EV-specific cargo composition is included to clarify why SC-EVs represent a particularly suitable extracellular vesicle source for neuropathy-associated oral wound repair. Finally, we evaluate current progress in scalable production, targeted engineering, and biomaterial-assisted delivery, while outlining remaining limitations related to standardization, potency assessment, long-term safety, and clinical translation. By integrating glial biology with extracellular vesicle nanomedicine, this review provides a focused framework for developing SC-EV-based therapeutics for diabetic oral mucosal lesions.

HSCR is caused by disruption of complex signaling pathways within the gene regulatory network (GRN) during enteric nervous system (ENS) development, including glial cell line-derived neurotrophic factor (GDNF) and GDNF family receptor alpha-1 (GFRα1). However, pathogenic variants in all GRN genes account for only ~ 80% cases; therefore, the epigenetic role remains to be elucidated. We compared GDNF and GFRα1 expression between HSCR patients and controls. qPCR revealed an upregulated GDNF expression in both ganglionic (206.37-fold) and aganglionic (126.35-fold) HSCR compared to control colons (ΔCT 7.65 ± 2.19 vs. 15.34 ± 1.34; p = 0.0001; and ΔCT 8.35 ± 2.57 vs. 15.34 ± 1.34; p = 0.0001). qPCR also showed an upregulated GFRα1 expression in both ganglionic (29.66-fold) and aganglionic (18.44-fold) HSCR compared to control colons (ΔCT 8.44 ± 1.98 vs. 13.33 ± 1.36; p = 0.0001; and ΔCT 9.13 ± 1.46 vs. 13.33 ± 1.36; p = 0.0001). In addition, no significant differences were observed between ganglionic and aganglionic segments for either GDNF or GFRα1 (ΔCT 7.65 ± 2.19 vs. 8.35 ± 2.57; p = 0.43; and ΔCT 8.44 ± 1.98 vs. 9.13 ± 1.46; p = 0.29). Our study demonstrates global aberrant expression of GDNF and GFRα1 in HSCR patients. These findings underscore the complexity of HSCR as a multifactorial disorder and highlight the importance of integrated signaling networks in ENS development, providing a rationale for further mechanistic and translational studies.

Delayed wound healing is a serious issue in diabetes, which is driven by metabolic and non-metabolic factors. Dysfunction and decreased lifespan of fibroblast cells play a key role in this condition. Lawsone (2-hydroxy-1,4-naphthoquinone), the main active component of Lawsonia inermis, is well known for its anticancer and regenerative effects, although these effects vary depending on concentration and experimental conditions. Regarding the importance of Diabetic wound treatment, this study examines how lawsone affects fibroblast behavior under high-glucose conditions, focusing on cell viability, apoptosis, intracellular Reactive Oxygen Species (ROS) levels, migration, and the expression of repair-related genes Nerve Growth Factor (NGF) and Transforming Growth Factor-beta (TGF-β). Human dermal fibroblasts were cultured under high-glucose (85-140mM) conditions, in the presence or absence of lawsone (10µg/mL). The MTT assay assessed cell viability, migration by the scratch assay, apoptosis by Annexin V- Fluorescein Isothiocyanate/Propidium Iodide (FITC/PI) flow cytometry, and ROS levels by 2',7'-dichlorodihydrofluorescein diacetate (H₂DCFDA) staining. quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) was used to quantify NGF and TGF-β mRNA expression. High glucose significantly impaired fibroblast viability and migration, increased apoptosis and ROS production, and downregulated TGF-β expression. Co-treatment with lawsone markedly restored cell viability and migration, suppressed oxidative stress and apoptosis, and enhanced NGF and TGF-β transcription, indicating reactivation of reparative pathways (p ≤ 0.05). Lawsone exhibits potent cytoprotective and pro-regenerative effects in human dermal fibroblasts under hyperglycemic stress, independent of its known anticancer activity. These findings highlight lawsone as a natural candidate for promoting diabetic wound healing and warrant further mechanistic and in vivo investigations.

This study aimed to investigate the effects of delivery mode and anesthesia type on fetal cord blood levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), which are neurotrophins involved in neuronal survival, differentiation, and stress adaptation during the perinatal period. In this cross-sectional, prospective study conducted from December 2023 to May 2024 at a single tertiary center, pregnant women were categorized based on delivery mode-vaginal delivery (VD) or cesarean section (CS). The CS group was subdivided according to anesthesia type: spinal or general. Cord blood NGF, BDNF, pH levels, and neonatal clinical outcomes were evaluated. Pregnant women with comorbidities and those receiving prenatal medical induction were excluded. The study included 150 pregnant women. Fifty of these patients had VD and 100 had CS. Sixty of the pregnant women who had a CS received spinal anesthesia, and 40 received general anesthesia. Cord blood NGF (54.20 ± 32.04 vs. 47.01 ± 27.28, p = 0.176) and BDNF (80.98 ± 41.94 vs. 70.08 ± 37.53, p = 0.123) values were not different in those who delivered with CS and VD. Cord blood NGF and BDNF values were higher in the spinal anesthesia group (p = 0.002, p = 0.003). The cord blood pH value was higher in the spinal anesthesia group than in the VD and general anesthesia groups (p = 0.004). Cord blood NGF and BDNF values were higher in the spinal anesthesia group. These findings support the hypothesis that regional anesthesia may be associated with reduced fetal stress and improved biochemical adaptation at birth.

BackgroundChronic pain affects over 30% of the global population and remains a major public health issue due to limited treatment efficacy and the need for mechanism-based, personalized approaches. Motor behavior is theorized to play a role in pain persistence through altered movement patterns, muscle recruitment, and proprioception. While motor behavior is linked to chronic pain, empirical evidence on underlying mechanisms, particularly cortical dynamics, remains scarce.ObjectiveThis study aimed to investigate longitudinal changes in cortical sensorimotor excitability and their relationship with maladaptive motor behaviors.MethodsThis prospective longitudinal study will follow 150 healthy participants, aged 18‐65 years, recruited from the community, experiencing experimentally induced muscle pain across 4 visits (day 0, day +2, day +4, and 5 days after pain resolution). Pain will be induced using intramuscular injections of nerve growth factor into the extensor carpi radialis brevis muscle. The primary outcome is motor variability assessed during multidimensional wrist movement tasks, and quantified using root-mean-square deviation and muscle synergies (derived from electromyographic recordings of 4 forearm muscles). Secondary outcomes include cortical mechanisms (electroencephalographic peak alpha frequency, transcranial magnetic stimulation mapping, short-interval intracortical inhibition, and intracortical facilitation), sensorimotor integration (evoked potential), cognitive control (multisource interference task), and endogenous pain modulation (conditioned pain modulation). Self-report questionnaires will assess pain intensity and disability (Patient-Rated Tennis Elbow Evaluation, McGill Pain Questionnaire–Short Form, and Likert Muscle Soreness Scale) as well as psychological factors such as fear of movement, pain-related beliefs, and coping strategies (Tampa Scale for Kinesiophobia, Pain Beliefs Questionnaire, and Coping Strategies Questionnaire–Revised). This sample size provides 80% power at 5% significance to detect a medium effect size across 23 predictor variables, with Bonferroni correction and 10% loss-to-follow-up allowance. Data will be analyzed using 2-level growth curve modeling to characterize interindividual differences in motor behavior, cortical dynamics, and pain processing trajectories.ResultsAll study procedures have been approved by the Western University Health Science Research Ethics Board (review reference 2025-125757-103291). Funding was provided by the Canadian Institutes of Health Research under grant number 517783 for the period 2024‐2029. Recruitment for the study began in April 2025, and all data collection is expected to be completed by 2028. As of April 2026, we have enrolled 26 participants. Results are expected to be published at the end of 2028.ConclusionsThe findings will advance understanding of motor behavior in pain and lay the foundation for personalized therapies, moving beyond current generic treatments that offer limited benefits.

Nerve guidance conduits (NGCs) hold considerable value in the field of nerve regeneration, yet current approaches display constraints regarding the diversity of loaded factors and their short sustained-release duration. Herein, a type of multifunctional inverse opal NGC integrating bone marrow mesenchymal stem cells (BMSCs) is presented for peripheral nerve repair. The generation of inverse opal scaffolds leverages the inversion of a monodisperse emulsion droplet template crafted by microfluidic technique. Ascribed to the biocompatibility and the cell-concentrating properties of the inverse opal scaffold, coupled with the inherent capabilities of BMSCs, such an NGC enables the secretion of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor, promoting the migration of rat Schwann cells and differentiation of pheochromocytoma 12 cells. Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) further revealed that three-dimensional (3D) BMSC spheroid culture in the inverse opal scaffold significantly upregulated bFGF, PDGF, and VEGF mRNA expression relative to two-dimensional (2D) culture. Further in vivo experimentation confirms the promising efficacy of NGCs loaded with BMSCs in repairing 10 mm sciatic nerve defects. These results underscore the substantial potential of this innovative design for peripheral nerve regeneration.

Deep endometriosis (DE) is the most severe subtype of endometriosis, marked by aggressive cellular behavior and debilitating pain. However, the molecular mechanisms underlying DE pathogenesis remain poorly understood. In this study, we identified interleukin (IL)-17A as a critical mediator to drive the pathological processes of DE. The level of IL-17A was elevated in DE tissues with T cells, mast cells, macrophages, and endometriosis stromal cells as sources of IL-17A. Functional assays demonstrated that IL-17A stimulates the proinflammatory cytokines such as IL-1β and IL-6 as well as enhances the proliferative and migratory capacities through activation of ERK1/2 and Notch1 signaling pathways. IHC staining further revealed levels of NICD and Ki67 are abundant and positively correlates with each other in DE lesions. In addition, PGP9.5+ nerves bundles are evidently detected in DE tissues as compared to normal endometria, pelvic endometriotic lesions, and ovarian endometrioma, which reflects the nature of severe pain in DE patients. Treatment with IL-17A induces the expression of nerve growth factor (NGF), a peptide growth factor to induce nerve infiltration. IHC staining reveals the significant positive correlation between PGP9.5+ nerves and NGF signals specifically in DE lesions. Collectively, these findings suggest that IL-17A promotes DE lesion progression by sustaining chronic inflammation and enhancing endometrial stromal cells proliferation, migration and nerve infiltration, thereby contributing to inflammation-associated neuropathic pain in affected patients.

BackgroundAdenomyosis is a major cause of chronic pelvic pain in women of reproductive age and significantly affects quality of life. Identifying biomarkers associated with pain mechanisms may improve understanding of disease pathophysiology and support the development of targeted therapeutic strategies. The mechanisms underlying adenomyosis-related pain are not fully understood but are thought to involve neurogenic factors such as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), which promote nerve fiber proliferation and sensitization. However, evidence regarding the relationship between BDNF, NGF, and pain severity in adenomyosis remains limited.ObjectiveTo compare BDNF and NGF levels in adenomyotic tissue, adjacent myometrium from patients with adenomyosis, and normal myometrium, and to evaluate their association with pain severity measured using the Visual Analog Scale (VAS).MethodsThis cross-sectional study analyzed BDNF and NGF levels in adenomyotic tissue, adjacent myometrium, and normal myometrium from 120 tissue samples. Neurotrophin levels were compared among tissue groups and evaluated for their association with pain severity using Visual Analogue Scale (VAS) scores.ResultsBDNF levels differed significantly among the three tissue groups (H = 99.364; p < 0.001), showing a decreasing trend from adenomyotic tissue to adjacent myometrium and then to normal myometrium. Post-hoc analysis confirmed significant differences across all pairwise comparisons (p < 0.001). NGF levels also differed significantly among groups (H = 96.056; p < 0.001), with significant differences in all pairwise comparisons (p < 0.001). No significant correlations were found between BDNF and VAS scores (ρ = −0.038; p = 0.817) or NGF and VAS scores (ρ = 0.125; p = 0.441).ConclusionAlthough BDNF and NGF levels differed significantly among tissue types, neither neurotrophin was significantly associated with pain severity, suggesting that additional mechanisms (eg, central sensitization or other inflammatory mediators) may underlie adenomyosis-related pain.

To observe the clinical therapeutic effect and effect mechanism of regulating-yinyang moxibustion therapy for primary dysmenorrhea (PD) of cold induced blood stasis, and conduct cost-effectiveness analysis. Sixty-six patients with PD of cold induced blood stasis were randomly divided into an observation group (33 cases, 1 case dropped out) and a control group (33 cases, 3 cases dropped out). The patients in the observation group received regulating-yinyang moxibustion therapy. Centered at Mingmen (GV4) and Guanyuan (CV4), moxibustion therapy for warming yang and benefiting kidney and for warming kidney and uterus was operated within an 8 cm to 10 cm radius separately. Moxibustion therapy for warming yang and benefiting kidney was delivered 10 days before menstruation (60 min), and that for warming kidney and uterus was performed 2 days before menstruation (40 min). One course of regulating-yinyang moxibustion therapy consisted of 1 operation for warming yang and benefiting kidney and 1 for warming kidney and uterus, and the treatment for 3 menstrual cycles was required. In the control group, gentle moxibustion therapy was operated at Guanyuan (CV4), Shenque(CV8), and bilateral Sanyinjiao (SP6), for 20 min at each acupoint. The intervention began 10 days before menstruation, once every other day; 1 course of treatment consisted of 5 interventions, and the treatment for 3 menstrual cycles was required.Before and after treatment, as well as in follow-up for 3 menstrual cycles after treatment, the scores of visual analogue scale(VAS) for pain, Cox menstrual symptom scale (CMSS), and traditional Chinese medicine (TCM) clinical syndromes were observed in the two groups. Before and after treatment, the contents of nerve growth factor (NGF) and prostaglandin F2α(PGF2α) in serum were detected. After treatment, the clinical therapeutic effect was compared, the cost-effectiveness analysis performed, and the sensitivity analysis conducted to verify the reliability of the results in the two groups. After treatment and in follow-up, the VAS scores for pain were reduced in comparison with those before treatment in each group (P<0.05), and the scores in the observation group were lower than those of the control group (P<0.05). The scores for the severity and duration in CMSS were lower than those before treatment in the observation group (P<0.05), and lower when compared with those in the control group (P<0.05). When compared with the score before treatment, the TCM syndrome scores in the observation group were reduced after treatment and in follow-up (P<0.05), and the score in the control group was lower after treatment (P<0.05). In follow-up, the TCM syndrome score in the observation group was lower than that in the control group (P<0.05). After treatment, the serum contents of NGF and PGF2α decreased in comparison with those before treatment (P<0.05), and the contents in the observation group were lower than those in the control group (P<0.05).After treatment, the total effective rate was 81.3% (26/32) in the observation group and was 73.3% (22/30) in the control group, without statistical significance (P>0.05). The cost-effectiveness analysis showed that the cost-effectiveness ratio(CER) in the observation group was lower than that in the control group. The sensitivity analysis found that there was no significant fluctuation of CER in either group. Regulating-yinyang moxibustion therapy effectively alleviates pain and TCM syndromes in patients with PD of cold induced blood stasis, which may be related to the decrease of the contents of NGF and PGF2α in serum. This therapy shows its advantages in terms of health economics compared with the conventional gentle moxibustion therapy.