Influenza Induces Lung Lymphangiogenesis Independent of YAP/TAZ Activity in Lymphatic Endothelial Cells.

The lymphatic system consists of a vessel network lined by specialized lymphatic endothelial cells (LECs) that are responsible for tissue fluid homeostasis and immune cell trafficking. The mechanisms for organ-specific LEC responses to environmental cues are not well understood. We found robust lymphangiogenesis during influenza A virus infection in the adult mouse lung. We show that the number of LECs increases 2-fold at 7 days post-influenza infection (dpi) and 3-fold at 21 dpi, and that lymphangiogenesis is preceded by lymphatic dilation. We also show that the expanded lymphatic network enhances fluid drainage to mediastinal lymph nodes. Using EdU labeling, we found that a significantly higher number of pulmonary LECs are proliferating at 7 dpi compared to LECs in homeostatic conditions. Lineage tracing during influenza indicates that new pulmonary LECs are derived from preexisting LECs rather than non-LEC progenitors. Lastly, using a conditional LEC-specific YAP/TAZ knockout model, we established that lymphangiogenesis, fluid transport and the immune response to influenza are independent of YAP/TAZ activity in LECs. These findings were unexpected, as they indicate that YAP/TAZ signaling is not crucial for these processes.

PHOTOTROPIN1 lysine 526 functions to enhance phototropism in Arabidopsis.

MAIN CONCLUSION: After blue-light exposure, ubiquitination of PHOTOTROPIN1 lysine 526 enhances phototropic responses. Arabidopsis blue-light photoreceptor, PHOTOTROPIN1 (PHOT1) mediates a series of blue-light responses that function to optimize photosynthesis efficiency. Blue-light sensing through the N-terminal sensory domain activates the C-terminal kinase activity of PHOT1, resulting in autophosphorylation. In addition to phosphorylation, PHOT1 lysine residue 526 (Lys526), after blue-light exposure, was found to carry a double glycine attachment, indicative of a possible ubiquitination modification. The functionality of PHOT1 Lys526 was investigated by reverse genetic approaches. Arginine replacements of PHOT1 Lys526, together with Lys527, complemented phot1-5 phot2-1 double mutant with attenuated phototropic bending, while blue-light responses: leaf expansion and stomatal opening, were restored to wild type levels. Transgenic seedlings were not different in protein levels of phot1 Lys526 527Arg than the wild type control, suggesting the reduced phototropic responses was not caused by reduction in protein levels. Treating the transformants with proteosome inhibitor, MG132, did not restore phototropic sensitivity. Both transgenic protein and wild type PHOT1 also had similar dark recovery of kinase activity, suggesting that phot1 Lys526 527Arg replacement did not affect the protein stability to cause the phenotype. Together, our results indicate that blocking Lys526 ubiquitination by arginine substitution may have caused the reduced phototropic phenotype. Therefore, the putative ubiquitination on Lys526 functions to enhance PHOT1-mediated phototropism, rather than targeting PHOT1 for proteolysis.

Fecal microbiota transplanted from old mice promotes more colonic inflammation, proliferation, and tumor formation in azoxymethane-treated A/J mice than microbiota originating from young mice.

Aging is a strong risk factor for colorectal cancer (CRC). It is well established that gut microbial dysbiosis can play a role in the etiology of CRC. Although the composition of the gut microbial community changes with age and is reported to become more pro-inflammatory, it is unclear whether such changes are also pro-tumorigenic for the colon. To address this gap, we conducted fecal microbiota transplants (FMT) from young (DY, ~6 wk) and old (DO, ~72 wk) donor mice into young (8 wk) recipient mice that were pre-treated with antibiotics. After initiating tumorigenesis with azoxymethane, recipients were maintained for 19 wk during which time they received monthly FMT boosters. Compared to recipients of young donors (RY), recipients of old donors (RO) had an approximately 3-fold higher prevalence of histologically confirmed colon tumors (15.8 vs 50%, Chi2 P = .03), approximately 2-fold higher proliferating colonocytes as well as significantly elevated colonic IL-6, IL-1ß and Tnf-α. Transcriptomics analysis of the colonic mucosa revealed a striking upregulation of mitochondria-related genes in the RO mice, a finding corroborated by increased mitochondrial abundance. Amongst the differences in fecal microbiome observed between DY and DO mice, the genera Ruminoclostridium, Lachnoclostridium and Marvinbryantia were more abundant in DY mice while the genera Bacteroides and Akkermansia were more abundant in DO mice. Amongst recipients, Ruminoclostridium and Lachnoclostridium were higher in RY mice while Bacteroides was higher in RO mice. Differences in fecal microbiota were observed between young and old mice, some of which persisted upon transplant into recipient mice. Recipients of old donors displayed significantly higher colonic proliferation, inflammation and tumor abundance compared to recipients of young donors. These findings support an etiological role for altered gut microbial communities in the increased risk for CRC with increasing age and establishes that such risk can be transmitted between individuals.

Genetic Factors Contribute to the Phenotypic Variability in GJB2-Related Hearing Impairment.

Recessive variants in GJB2 are the most important genetic cause of sensorineural hearing impairment (SNHI) worldwide. Phenotypes vary significantly in GJB2-related SNHI, even in patients with identical variants. For instance, patients homozygous for the GJB2 p.V37I variant, which is highly prevalent in the Asian populations, usually present with mild-to-moderate SNHI; yet severe-to-profound SNHI is occasionally observed in approximately 10% of p.V37I homozygotes. To investigate the genomic underpinnings of the phenotypic variability, we performed next-generation sequencing of GJB2 and other deafness genes in 63 p.V37I homozygotes with extreme phenotypic severities. We identified additional pathogenic variants of other deafness genes in 5 of the 35 patients with severe-to-profound SNHI. Furthermore, we conducted case-control association analyses for 30 unrelated p.V37I homozygotes with severe-to-profound SNHI against 28 p.V37I homozygotes with mild-to-moderate SNHI, and 120 population controls from the Taiwan Biobank. We found that the severe-to-profound group had a higher frequency of the crystallin lambda 1 (CRYL1) variant (rs14236), located upstream of GJB2, than the mild-to-moderate and Taiwan Biobank groups. Our results demonstrated that pathogenic variants in other deafness genes and a possible modifier, the CRYL1 rs14236 variant, may contribute to phenotypic variability in GJB2-realted SNHI, highlighting the importance of comprehensive genomic surveys to delineate the genotype-phenotype correlations.

Identification of nine novel variants across PAX3, SOX10, EDNRB, and MITF genes in Waardenburg syndrome with next-generation sequencing.

BACKGROUND: Waardenburg syndrome (WS) is a hereditary, genetically heterogeneous disorder characterized by variable presentations of sensorineural hearing impairment and pigmentation anomalies. This study aimed to investigate the clinical features of WS in detail and determine the genetic causes of patients with clinically suspected WS. METHODS: A total of 24 patients from 21 Han-Taiwanese families were enrolled and underwent comprehensive physical and audiological examinations. We applied targeted next-generation sequencing (NGS) to investigate the potential causative variants in these patients and further validated the candidate variants through Sanger sequencing. RESULTS: We identified 19 causative variants of WS in our cohort. Of these variants, nine were novel and discovered in PAX3, SOX10, EDNRB, and MITF genes, including missense, nonsense, deletion, and splice site variants. Several patients presented with skeletal deformities, hypotonia, megacolon, and neurological disorders that were rarely seen in WS. CONCLUSION: This study revealed highly phenotypic variability in Taiwanese WS patients and demonstrated that targeted NGS allowed us to clarify the genetic diagnosis and extend the genetic variant spectrum of WS.

Physiological Impact of a Synthetic Elastic Protein in Arterial Diseases Related to Alterations of Elastic Fibers: Effect on the Aorta of Elastin-Haploinsufficient Male and Female Mice.

Elastic fibers, made of elastin (90%) and fibrillin-rich microfibrils (10%), are the key extracellular components, which endow the arteries with elasticity. The alteration of elastic fibers leads to cardiovascular dysfunctions, as observed in elastin haploinsufficiency in mice (Eln+/-) or humans (supravalvular aortic stenosis or Williams-Beuren syndrome). In Eln+/+ and Eln+/- mice, we evaluated (arteriography, histology, qPCR, Western blots and cell cultures) the beneficial impact of treatment with a synthetic elastic protein (SEP), mimicking several domains of tropoelastin, the precursor of elastin, including hydrophobic elasticity-related domains and binding sites for elastin receptors. In the aorta or cultured aortic smooth muscle cells from these animals, SEP treatment induced a synthesis of elastin and fibrillin-1, a thickening of the aortic elastic lamellae, a decrease in wall stiffness and/or a strong trend toward a reduction in the elastic lamella disruptions in Eln+/- mice. SEP also modified collagen conformation and transcript expressions, enhanced the aorta constrictive response to phenylephrine in several animal groups, and, in female Eln+/- mice, it restored the normal vasodilatory response to acetylcholine. SEP should now be considered as a biomimetic molecule with an interesting potential for future treatments of elastin-deficient patients with altered arterial structure/function.

Hearing Features and Cochlear Implantation Outcomes in Patients With Pathogenic MYO15A Variants: a Multicenter Observational Study.

OBJECTIVES: Recessive variants in the MYO15A gene constitute an important cause of sensorineural hearing impairment (SNHI). However, the clinical features of MYO15A-related SNHI have not been systemically investigated. This study aimed to delineate the hearing features and outcomes in patients with pathogenic MYO15A variants. DESIGN: This study recruited 40 patients with biallelic MYO15A variants from 31 unrelated families. The patients were grouped based on the presence of N-terminal domain variants (N variants). The longitudinal audiological data and for those undergoing cochlear implantation, the auditory and speech performance with cochlear implants, were ascertained and compared between patients with different genotypes. RESULTS: At the first audiometric examination, 32 patients (80.0%) presented with severe to profound SNHI. Patients with at least one allele of the N variant exhibited significantly better hearing levels than those with biallelic non-N variants (78.2 ± 23.9 dBHL and 94.7 ± 22.8 dBHL, respectively) (p = 0.033). Progressive SNHI was observed in 82.4% of patients with non-profound SNHI, in whom the average progression rate of hearing loss was 6.3 ± 4.8 dBHL/year irrespective of the genotypes. Most of the 25 patients who underwent cochlear implantation exhibited favorable auditory and speech performances post-implantation. CONCLUSIONS: The hearing features of patients with biallelic pathogenic MYO15A variants are characterized by severe to profound SNHI, rapid hearing progression, and favorable outcomes with cochlear implants. Periodic auditory monitoring is warranted for these patients to enable early intervention.

Room-temperature multiple ligands-tailored SnO2 quantum dots endow in situ dual-interface binding for upscaling efficient perovskite photovoltaics with high VOC.

The benchmark tin oxide (SnO2) electron transporting layers (ETLs) have enabled remarkable progress in planar perovskite solar cell (PSCs). However, the energy loss is still a challenge due to the lack of "hidden interface" control. We report a novel ligand-tailored ultrafine SnO2 quantum dots (QDs) via a facile rapid room temperature synthesis. Importantly, the ligand-tailored SnO2 QDs ETL with multi-functional terminal groups in situ refines the buried interfaces with both the perovskite and transparent electrode via enhanced interface binding and perovskite passivation. These novel ETLs induce synergistic effects of physical and chemical interfacial modulation and preferred perovskite crystallization-directing, delivering reduced interface defects, suppressed non-radiative recombination and elongated charge carrier lifetime. Power conversion efficiency (PCE) of 23.02% (0.04 cm2) and 21.6% (0.98 cm2, VOC loss: 0.336 V) have been achieved for the blade-coated PSCs (1.54 eV Eg) with our new ETLs, representing a record for SnO2 based blade-coated PSCs. Moreover, a substantially enhanced PCE (VOC) from 20.4% (1.15 V) to 22.8% (1.24 V, 90 mV higher VOC, 0.04 cm2 device) in the blade-coated 1.61 eV PSCs system, via replacing the benchmark commercial colloidal SnO2 with our new ETLs.

Hearing Impairment with Monoallelic GJB2 Variants: A GJB2 Cause or Non-GJB2 Cause?

Recessive variants in GJB2 are the most common genetic cause of sensorineural hearing impairment. However, in many patients, only one variant in the GJB2 coding region is identified using conventional sequencing strategy (eg, Sanger sequencing), resulting in nonconfirmative diagnosis. Conceivably, there might be other unidentified pathogenic variants in the noncoding region of GJB2 or other deafness-causing genes in these patients. To address this, a next-generation sequencing-based diagnostic panel targeting the entire GJB2 gene and the coding regions of 158 other known deafness-causing genes was designed and applied to 95 patients with nonsyndromic sensorineural hearing impairment (including 81 Han Taiwanese and 14 Mongolian patients) in whom only a single GJB2 variant had been detected using conventional Sanger sequencing. The panel confirmed the genetic diagnosis in 24 patients (25.3%). Twenty-two of them had causative variants in several deafness-causing genes other than GJB2, including MYO15A, MYO7A, TECTA, POU4F3, KCNQ4, SLC26A4, OTOF, MT-RNR1, MITF, WFS1, and USH2A. The other two patients had causative variants in GJB2, including a Taiwanese patient with a mosaic maternal uniparental disomy c.235delC variant (approximately 69% mosaicism) and a Mongolian patient with compound heterozygous c.35dupG and c.35delG variants, which occurred at the same site. This study demonstrates the utility of next-generation sequencing in clarifying the genetic diagnosis of hearing-impaired patients with nonconfirmative GJB2 genotypes on conventional genetic examinations.

A rare complication of cerebral venous thrombosis during simple percutaneous coronary intervention: A case report.

RATIONALE: Cerebrovascular accidents (CVAs) after percutaneous coronary intervention (PCI), although rare, are associated with high in-hospital morbidity and mortality rates. Cerebral venous thrombosis (CVT) is an uncommon cause of CVAs compared with arterial disease but is associated with favorable outcomes in most cases. We present a rare case of CVT following a simple PCI procedure with stent implantation, which has not been previously reported in the literature. PATIENT CONCERNS: A 78-year-old woman with hypertension, hyperlipidemia, and coronary artery disease received simple PCI with stent implantation. After PCI, she developed a throbbing headache with nausea and vomiting, with her blood pressure increasing to 190/100 mmHg. Drowsiness, disorientation, and neck stiffness were noted. Neurological complication due to the PCI procedure was highly suspected. DIAGNOSIS: Noncontrast brain computed tomography was performed along with emergency neurological consultation, and the patient was diagnosed as having acute CVT. INTERVENTIONS: The patient was treated with anti-intracranial pressure therapy and anticoagulation therapy through low-molecular-weight heparin and was subsequently treated with warfarin. OUTCOMES: After treatment, the patient's symptoms and signs gradually subsided, and her clinical condition improved. She was discharged with full recovery thereafter. LESSONS: A case of acute CVT, a rare, and atypical manifestation of venous thromboembolism and CVA, complicated simple PCI with stent implantation. During PCI, identifying patients with a high risk of a CVA is critical, and special care should be taken to prevent this devastating complication.

Resisting the racialization of medical deservingness: How Latinx nurses produce symbolic resources for Latinx immigrants in clinical encounters.

This paper investigates how Latinx nurses resisted the racialization of medical un-deservingness against co-ethnic immigrants in everyday clinical encounters. Drawing on 26 in-depth interviews and dialoguing with the literature on minority professionals, we find that, as a form of racialized equity work, Latinx nurses produced certain symbolic resources, specifically the interactional signals to counteract Latinx patients' internalization of un-deservingness and other medical staff's open hostility towards these "undeserving illegals." Latinx nurses hybridized neoliberal norms (self-sufficiency and responsibility) and social justice values (including healthcare as a universal right and compassion for members of the community): they emphasized Latinx immigrants' efforts at "becoming" self-sufficient and clinically responsible, debunked the relevance of citizenship to a right to healthcare, and highlighted their communal bonds with co-ethnic patients. Meanwhile, accentuating these communal bonds revealed hefty loads of previously self-censored healthcare needs among Latinx patients, which compelled Latinx nurses to reassert some professional boundaries. Whereas some Latinx nurses were able to engage in "moralized boundary-drawing," others experienced setting professional boundaries as "demoralizing boundary-drawing," which resulted in burnout, disillusionment, or internalized racism. Our findings indicate that the path to de-racializing medical deservingness needs to be multi-tiered. Latinx nurses' racialized equity work of generating symbolic resources for Latinx immigrants is only sustainable if supported by non-Latinx colleagues' cross-ethnic equity work. Furthermore, everyday resistance in clinical encounters is necessarily incomplete unless state-level policy initiatives transform the currency of symbolic capital for medical deservingness.

The "Societalization" of pandemic unpreparedness: lessons from Taiwan's COVID response.

Adopting a Civil Sphere Theory framework, we argue that Taiwan's efforts at containing COVID-19 resulted from its "societalization" of pandemic unpreparedness, which was triggered by the 2003 SARS outbreak and resumed during the COVID-19 pandemic. Societalization refers to the process through which institutional failures are transformed into societal crises, with the civil sphere mobilized to discuss institutional dysfunctions, push for reforms, and attempt to democratize or otherwise transform institutional cultures. The societalization of pandemic unpreparedness in Taiwan led to reforms of the public health administration and the medical profession, thereby establishing state mechanisms for encouraging early responses and coordinating centralized command during outbreaks, and healthcare infrastructures for coordinating patient transfer and ensuring supplies of personal protective equipment. Reflections upon past uncivil acts among citizens motivated the civil sphere to foster a discourse of interdependence, redefining the boundaries between individual choices and civic virtues. Meanwhile, unaddressed challenges remained, including threats related to Taiwan's political polarization. Our paper challenges the thesis of "authoritarian advantage," highlighting how democratic societies can foster social preparedness to respond to crises. By illustrating how societalization can reach temporary closures but become reactivated subsequently, our study extends the theory of societalization by explicating its historical dimension.

Charge Energetics and Electronic Level Changes At the Copper(II) Phthalocyanine/Fullerene Junction Upon Photoexcitation.

Energy offset at the donor (D)/acceptor (A) interface plays an important role in charge separation in organic photovoltaics. Its magnitude determines the charge separation process under illumination. Extensive studies have been carried out for elucidating the charge transfer (CT) process at different D/A junctions. These works lead to two different views: upon photoexcitation, energies would be (1) consumed in molecular polarization and orientation such that those opposite charges would overcome mutual Coulombic attractive potential at the interface and (2) spent on promoting charges to high-lying delocalized energy states (i.e., hot states), which is necessarily important prior to charge separation. Under these two scheme of studies, the electronic structures and the charge behaviors at the D/A interface should be different under photoexcitation, yet there is so far no direct experimental approach for probing the changes in electronics structures (i.e., Fermi level, vacuum level, frontier molecular orbitals, etc.) upon photoexcitation. Herein, a modified photoelectron spectroscopy (PES) system with an additional solar simulator is used to study the charge distributions and electronic interactions for a standard D/A heterojunction (i.e., copper phthalocyanine (CuPc)/ fullerene (C60)) under photoexcitation. CT states formed as a result of photon energy transfer at the CuPc/C60 junction. Subsequent superpositions of charge transfer and electron polarization effects increase the D/A energy level offsets from 0.75 (ground state measured in the dark) to 1.07 eV (high-lying state measured upon illumination). We showed that there is excess energy consumed for a subtle change in the energy level alignment of the CuPc/C60 junction under illumination, suggesting a new insight for the energy loss mechanism during the photocharge generation processes.

Organic-Inorganic Charge Transfer Complex with Charge Modulation after Electrical Pre-biasing.

Several breakthroughs in organic optoelectronic devices with new applications and performance improvement have been made recently by exploiting novel properties of charge transfer complexes (CTCs). In this work, a CTC film formed by coevaporating molybdenum(VI) oxide and pentacene (MoO3:pentacene) shows a strong dipole of 2.4 eV with direction controllability via pre-biasing with an external voltage. While CTCs are most widely known for their much red-shifted energy gaps, there is so far no report on their controllable dipoles. By controlling this dipole with an electrical pre-bias in a MoO3:pentacene CTC based device, current changes over 2 orders of magnitude can be achieved. Kelvin probe force microscopy further confirms that surface potential of the MoO3:pentacene film can be modulated by an external electric field. It is shown for the first time that a dipole of controllable direction can be set up inside a CTC layer by pre-biasing. This concept is further tested by incorporating the CTC layer in organic photovoltaic (OPV) devices. It was demonstrated that by pre-biasing the OPV devices in different directions, their open circuit voltages (Voc) can be significantly tuned via the built-in potentials.

Black Bean Anthocyanin-Rich Extract from Supercritical and Pressurized Extraction Increased In Vitro Antidiabetic Potential, While Having Similar Storage Stability.

Black bean is a source of anthocyanins and other phenolic compounds that are associated with health benefits. This work aimed to optimize the extraction and determine the stability and biological potential of black bean anthocyanin-rich extracts recovered by supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE). The highest concentration of anthocyanins and total phenolic compounds were recovered with SFE using 300 bar, 60 °C and co-solvent ethanol/distilled water (50/50, v/v). Eleven non-colored phenolic compounds were identified in SFE extract using Ultra performance liquid chromatography - Electrospray ionization-Quadrupole -Time of flight - Mass spectrometry (UPLC-ESI-QToF-MS/MS). Myricetin, syringic acid, rutin hydrate and chlorogenic acid presented the highest relative area among identified compounds. Compared to leaching extraction, SFE extracts showed a similar storage stability at 4, 25 and 32 °C (p < 0.05), but with a higher antioxidant potential (2,2-diphenyl-1-picryl-hydrazil (DPPH) IC50: 0.078 ± 0.01; 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) IC50: 0.161 ± 0.03) and antidiabetic potential (α-amylase IC50: 124.76 ± 12.97; α-glucosidase IC50: 31.30 ± 0.84; dipeptidyl peptidase-IV IC50: 0.195 ± 0.01). SFE extraction is an efficient method to obtain anthocyanins and other phenolic compounds with exceptional biological potential.

Effects of Hydrogen Bonds between Polymeric Hole-Transporting Material and Organic Cation Spacer on Morphology of Quasi-Two-Dimensional Perovskite Grains and Their Performance in Light-Emitting Diodes.

Perovskite is emerging as a novel emitter in solution-processed light-emitting diodes (LEDs). In these LEDs, morphology, especially the grain size of perovskite, plays a key role in determining electroluminescence performance. Several studies have shown that sizes of the perovskite grains can be controlled by the contact angle between the perovskite solution and the substrate. In this work, we found that in the quasi-two-dimensional (2D) system, the perovskite grain size can be substantially refined when there are hydrogen bonding between the perovskite's organic spacer and the substrates. In fact, for quasi-2D perovskite, with the presence of such hydrogen bond, its effects on the perovskite grain size overshadow the contact angle's effect. We demonstrated that perovskite with refined grains can form amine- or carbazole-based polymers which can form N···H hydrogen bonding with the perovskite's organic spacer. Using these polymers as hole-transporting layers on poly(3,4-ethylenedioxythiophene):polystyrene sulfonate, external quantum efficiency of CsPbBr3-based LEDs can be enhanced from 1.5 to 10.0% without passivation treatment. This work suggests that bonding between perovskite precursors and the substrate can have significant influence on the morphology of the final perovskite grains and their optoelectronic performance.

The effect of electroacupuncture merged with rehabilitation for frozen shoulder syndrome: A single-blind randomized sham-acupuncture controlled study.

PURPOSE: Frozen shoulder syndrome (FSS) causes pain and reduces the range of motion in the shoulder joint. To investigate the short and medium-term effects of electroacupuncture in people with FSS, we evaluated the therapeutic effects of true and sham electroacupuncture on pain relief and improvement of shoulder function. METHODS: In this randomized, single-blind controlled clinical trial, 21 subjects with FSS were randomly assigned to two groups: a true electroacupuncture group (TEAG) and a sham electroacupuncture group (SEAG). The two groups underwent 18 sessions of treatment over approximately 6-9 weeks and were then followed up at 1, 3, and 6 months. Their effectiveness for alleviating the intensity of shoulder pain was evaluated with a visual analog scale (VAS), while improved shoulder mobility was evaluated by the active range of motion (AROM) and passive range of motion (PROM), and shoulder functional ability was evaluated using the Shoulder Pain and Disability Index (SPADI). RESULTS: It demonstrated that the TEAG or SEAG showed lasting effects at 1, 3, and 6 months, although with no significant difference between these two groups in the shoulder functional ability outcomes. However, the decline in the VAS occurred earlier in the TEAG than the SEAG. Also, there was much more improvement in AROM for flexion and abduction in the TEAG than the SEAG. An increase in the abduction angle after electroacupuncture and manual rehabilitation was also apparent. CONCLUSION: These results suggest that electroacupuncture plus rehabilitation may provide earlier pain relief for patients with FSS and could be applied clinically.