Androgen signaling comprises nongenomic and genomic pathways. Nongenomic actions are not related to the binding of the androgen receptor (AR) and occur rapidly. The genomic effects implicate the binding to a cytosolic AR, leading to protein synthesis. Both events are independent of each other. Genomic effects have been associated with different pathologies such as vascular ischemia, hypertension, asthma, and cardiovascular diseases. Catecholamines play a crucial role in regulating vascular smooth muscle (VSM), airway smooth muscle (ASM), and cardiac muscle (CM) function and tone. Body. Testosterone (T) favors vasoconstriction, and its concentration fluctuation during life stages can affect the vascular tone and might contribute to the development of hypertension. In the VSM, T increases α1-adrenergic receptors (α1-ARs) and decreases adenylyl cyclase expression, favoring high blood pressure and hypertension. Androgens have also been associated with asthma. During puberty, girls are more susceptible to present asthma symptoms than boys because of the increment in the plasmatic concentrations of T in young men. In the ASM, β2-ARs are responsible for the bronchodilator effect, and T augments the expression of β2-ARs evoking an increase in the relaxing response to salbutamol. The levels of T are also associated with an increment in atherosclerosis and cardiovascular risk. In the CM, activation of α1A-ARs and β2-ARs increases the ionotropic activity, leading to the development of contraction, and T upregulates the expression of both receptors and improves the myocardial performance.  The aim of this review is an updated analysis of the role of androgens in the adrenergic system of vascular, airway, and cardiac myocytes.  Androgens play an essential role in the adrenergic system of vascular, airway, and cardiac myocytes, favoring either a state of health or disease. While the use of androgens as a therapeutic tool for treating asthma symptoms or heart disease is proposed, the vascular system is warmly affected. Afterward, 17α-hydroxypregnenolone is converted to dehydroepiandrosterone (DHEA) by cytochrome P450c17/CYP17A1 .β-HSD are expressed in cardiac i) and the activation of protein kinase C (PKC) i and induce myocardial contraction i and favoring muscle contraction i i . Vasocong others –174. Thegh VDCCs . In the  rapidly , 177.2+. The influx of this ion is also carried out by nonselective cation channels such as receptor-operated Ca2+ channels (ROCCs), store-operated Ca2+ channels (SOCCs), and transient receptor potential (TRP) channels. The Ca2+ influx exerted by these channels is thought to be triggered by agonists such as norepinephrine, vasopressin, and acetylcholine via GPCRs linked to the phospholipase Cβ (PLCβ) signaling pathway and the formation of IP3 and DAG. This last second messenger regulates the activity of ROCCs, and IP3 induces depletion of internal Ca2+ stores leading to capacitative Ca2+ entry through SOCCs i in the ASM cells. The mechanisms responsible for the Ca2+ influx are carried out by transient receptor potential canonical 3 (TRPC3), L-VDCCs and T-VDCCs, ROCCs, SOCCs, and reverse-mode Na+/Ca2+ exchanger (NCXREV) i is restored by two ATPases: sarcoplasmic reticulum Ca2+-ATPase (SERCA) and plasma membrane Ca2+-ATPase (PMCA) [β2-AR is the main adrenoceptor subtype responsible for the bronchodilator effect [2+ availability and inactivating myosin light-chain kinase [β2-AR favors hyperpolarization and relaxation of the ASM through the opening of different K+ channels [+ channels are the Ca2+-activated K+ channels (KCa) and KV [Ca are activated by increases in [Ca2+]i and through the cAMP-PKA signaling pathway i and the muscular tone through the inhibition of L-VDCCs and TRPC3 i determines the contractile function of the heart through distinct Ca2+-handling proteins. In the sinoatrial node, the pacemaker cells start depolarization of the cardiac myocytes. This process is regulated by the parasympathetic nervous system (PNS) and the SNS i and the formation of the Ca2+-calmodulin complex with the further activation of MLCK. Afterward, Ca2+ is sequestered to the SR by SERCA, and the cell takes it out by the Na+-Ca2+ exchanger in its forward mode (NCX). In addition, KCa channels are activated, leading to membrane hyperpolarization. These are the main mechanisms responsible for CM relaxation i i , 255. PK rapidly . In huma effects . The β2-ed by Gs .2+-handling proteins. Treatment of rat ventricular cardiomyocytes with T increased the levels of gene expression of L-VDCC, β1-AR, and NCX with 8 and 24 hours of exposure i [b[Ca2+]i, via the upregulation of the Ca2+ channels aforementioned, might contribute to chronic cardiac pathogenesis when T levels are elevated.Besides their electrophysiological properties, VDCCs are also classified using a standard nomenclature based on their molecular features of the pore-forming 1–CaV3.3 . In thisb[Ca2+]i , 262. Th2+ handling and the cardiac contraction/relaxing outcomes. On the one hand, a group of researchers demonstrated in cultured rat cardiomyocytes that the acute exposure to T rapidly increased [Ca2+]i, and this augment was not abolished by an antagonist of the androgen receptor. Elegantly, the authors confirmed that the mechanism involved in the T-induced increase in [Ca2+]i was mediated by the activation of a plasma membrane AR associated with a pertussis toxin- (PTX-) sensitive G protein (Gi/o) and with the activation of the PLC-IP3 signaling pathway leading to cardiac hypertrophy and failure [βγ-subunits of the Gi/o proteins [While the nongenomic effects of T on the CM have been reported, the studies are scarce and seem to be contradictory regarding the Ca failure . The actproteins , 265. Onproteins , 266.Gender-related differences in cardiovascular disease (CVD) seem to be affected by age. It is well documented that the risk of dying for men between ages 45 and 64 from a CVD is higher than for women in the premenopausal period; however, there is a slight increase in the risk of CVD death in women after menopause .The role of sex hormones in CVD is still unclear; particularly, a controversy about the effects of T in cardiovascular (CV) health and disease currently exists in the medical community. It is generally accepted that normal levels of T are beneficial for CV health in men, and a decline in these levels is related to an increase in CV events . Neverthα1-AR signaling pathway. The activation of α1-AR improves the myocardial performance after an infarction, reducing injury and arrhythmias [It has been proposed that low plasmatic levels of T in older men are associated with an increase in atherosclerosis and cardiovascular risk, suggesting that this androgen plays a cardioprotective role against CVDs, such as coronary heart disease (CHD) and chronic heart failure (CHF) . Moreovehythmias . CHD is hythmias . In thishythmias , 292. Fuhythmias . These ohythmias .α- and β-ARs controlling the CM tone. However, during myocardial ischemia, the release of norepinephrine increases the risk and contributes to cardiac injury [β1-AR triggered proapoptotic pathways, weakening the cardiac structure and accelerating heart injury and failure progression [The SNS (through norepinephrine) activates c injury , 296. Ingression , 297. Thgression .α1-ARs may play an important role in cardioprotection, specifically, the α1A-subtype. The overexpression of α1A-AR can improve the outcome after myocardial infarction [α1-AR expression and augmented the cardiac responses, leading to a reduction in ischemia and cardiac injury [α1- and β1-AR in perfused rat hearts  through the protection of the cardiac β-adrenergic system. Notably, the stimulation of the AR by T upregulated the expression of β2-AR, improving the myocardial performance , an enzyme implicated in β1-AR and β2-AR dysregulation in CHF [α2-AR/catecholamine production axis [β-AR responsiveness and diminishes the activity of GRK2 [β1-AR expression in the heart would be directly influenced by anabolic-androgenic steroids  [β and TNF-α and plasmatic levels of total cholesterol [β1- and β2-AR in the cardiac right atrium, provoked the prolongation of the QTc interval, and increased the BP [β1-AR protein expression in the left ventricle [The genomic regulation of the  failure , 312. Inn in CHF –315. Morion axis . Also, e of GRK2 . Intereses of T) . The usees of T) , 319. Hoes of T) –322. Forlesterol . Furtherlesterol . In thisd the BP . In addientricle . These dT, as a treatment in CVD, should only be considered for male patients with a diagnosis of hypogonadism. Moreover, due to the increase of T therapy for postmenopausal women, the potential risk of developing CDV events needs further research [Given the differences between studies showing the protective role of T in CV events and reports pointing out adverse CVD outcomes, it has been remarkably proposed that the use of research , 275.α1-AR expression, leading to high BP and hypertension. Moreover, in the airway smooth muscle, T, via nongenomic action, potentiates the β-adrenergic-induced relaxation through the inhibition of COMT or by the abolition of extraneuronal uptake. This androgen, via a genomic effect, also augments the expression of β2-AR and induces an increase in the relaxing responses to salbutamol. In the cardiac muscle, T upregulates the expression of α1A-AR and β2-AR mediated by the AR signaling, improving the myocardial performance. Moreover, T also increments β1-AR expression, improving the cardiomyocytes' function; however, the enhancement in muscle work during a long period ends up developing hypertrophy and heart failure.The adrenergic system plays a pivotal role in the control of vascular, airway, and cardiac physiology. A relationship between androgens with the adrenergic system of these tissues is proposed. This review summarizes that, in the vascular smooth muscle, T, via the androgen receptor, reduces the AC expression and increases the Consequently, we might argue that androgen genomic actions have deleterious effects in the VSM favoring hypertension. Nevertheless, in the ASM, nongenomic and genomic actions of androgens contribute to diminish the hyperresponsiveness of this tissue, favoring the absence of asthma symptoms. Therefore, androgens could be a therapeutic alternative for asthma treatment. However, in heart diseases, further research is required to determine the possible therapeutic use of androgens in these ailments.β-DHT, a well-known T metabolite without genomic effects, could be a prospective therapeutic agent for the treatment of these illnesses.Finally, the use of T and DHEA as a therapeutic tool for the treatment of asthma symptoms or some cardiovascular diseases, is questionable. T has virilizing adverse effects, androgenic actions that favor prostate cancer, and its aromatization leads to the production of estrogens. Additionally, DHEA is further biotransformed into various sex steroids, such as T and estrogens, with their subsequent effects. However, 5


Changes in β-AR responsiveness are thought to be an important factor that contributes to the diabetic cardiac dysfunction. Diabetes related changes in β-AR expression, signaling, and β-AR mediated cardiac function have been studied by several investigators for many years. In the present review, we have screened PubMed database to obtain relevant articles on this topic. Our search has ended up with wide range of different findings about the effect of diabetes on β-AR mediated changes both in molecular and functional level. Considering these inconsistent findings, the effect of diabetes on cardiac β-AR still remains to be clarified.Diabetes is a chronic, endocrine disorder that effects millions of people worldwide. Cardiovascular complications are the major cause of diabetes-related morbidity and mortality. Cardiac β Diabetes is an endocrine disorder due to partial or complete insulin deficiency or to insulin resistance in the target tissues. According to the World Health Organization (WHO) the number of diabetic individuals rose from 108 million in 1980 to 422 million in 2014 i transient amplitude were increased when murine cardiomyocytes were treated with PTX -CGP 12,177, [3H]-dihydroalprenolol ([3H]-DHA), [125I]-iodocyanopindolol and [125I]-iodopindolol have been used as ligands for detection of β-AR. In most of these studies, except one -DHA binding 8 weeks after STZ injection. Such, downregulation of β-AR has been confirmed in many studies by using different methods at both protein and mRNA level  mice i change in response to isoprenaline was reported in diabetic cells [β-AR mediated responses have been also evaluated by the change in Catic rats . Isoprentic rats . Paralleic cells .There are relatively fewer studies which have investigated β-AR mediated in vivo in T1DM. In vivo cardiac parameters such as rate of contraction/relaxation and heart rate were attenuated after isoprenaline treatment in STZ diabetic mice . SimilarAs mentioned in the previous sections, T2DM animal models have received attention because of their greater translational value. Parallel to the studies in T1DM models, mostly rats, but also mice have been used in studies on T2DM. Isoprenaline-stimulated sarcomere shortening in cardiomyocytes isolated from high fat fed mice was not found different compared to control cardiomyocytes . IsoprenThe inotropic effect of isoprenaline was not altered in left ventricular trabecular muscle in neonatal non-insulin-dependent diabetes model . Similar1-AR is the main subtype which regulates heart function in both healthy and diabetic rats whereas β2-AR has an indirect influence on β-AR mediated responses in the diabetic heart [β-AR mediated response on contraction and relaxation was impaired in ZDF rats by using the Langendorff heart preparation. It has been suggested that βic heart . Dobutamic heart . Impaireic heart . Left veic heart .1-AR is the main subtype to modulate chronotropic effect in control and diabetic animals [The in vivo effect of isoprenaline was attenuated in high fat fed rats ,112,115. animals . Further animals .Cardiovascular complications are the major risk factor for mortality in individuals with diabetes . It is i1- and β2-AR and less if any in β3-AR as explained in 1-AR and β3-AR was found to be decreased and increased respectively, consistent with the protein level. However, it was found that the β2-AR mRNA level was increased despite the reduced β2-AR protein level, and this has been referred to the sensitization of the receptor [3-AR is one of the most consistent findings in the STZ model of T1DM. Similarly, upregulation of β3-ARs have been reported in the heart failure. In both human [3-ARs was found to be increased. On the other hand, these studies have indicated conflicting results about the β3-AR mediated relaxation in the failed heart. The response to β3-AR stimulation was blunted in the human heart [3-ARs have been also related to a protective role in cardiac ischemia and reperfusion injury. The beneficial effects due to β3-AR stimulation have been demonstrated in this pathology [3-AR stimulation in ischemia and reperfusion injury was seen only in control rats not in high fat high sucrose (HFS) fed mice [3-ARs in HFS mice. These findings imply that the expressional or functional changes of β3-ARs may contribute to the cardiac pathologies. Most investigators have reported a decreased expression of β-AR by using radioligand binding studies in rodent models of T1DM, but a smaller number of studies did not confirm such changes, and an increased expression has been reported in two studies from one group of investigators ,80. The receptor . While nth human  and canith human , the expan heart  while itan heart . It has an heart . β3-ARs athology ,184. Of fed mice . This reCardiac β-AR post-receptor signaling pathway regulation may contribute to changes in β-AR responsiveness in diabetes. Supporting that, reduced β-AR responsiveness has been reported despite unchanged β-AR protein and/or mRNA level in diabetic state in some studies ,115. HowSame as the molecular changes in β-AR, in vivo and in vitro studies have yielded different results. Increased and decreased response to the same agonist, in the whole heart  and in tIn conclusion, since the late 1970s, diabetes related changes in cardiac β-AR have attracted attention of several researchers but their findings are controversial. At the molecular level, antibody-based approaches are troubled by low target selectivity and in general studies have a very poor reproducibility rate. The studies primarily have been conducted on rodent cardiac tissues and few have been done in other species including humans. In preclinical studies, mostly male animals have been preferred, there are relatively fewer studies which have been done in female animals. Even though 90% of diabetic patients have T2DM , more st


Background: Androgen receptor (AR) has been described to play a prominent role in male breast cancer (MBC). It maps on chromosome X, and recent reports indicate that X-chromosome polysomy is frequent in MBC. Since the response to anti-androgen therapy may depend on AR polysomy and on its overexpression similarly to prostate cancer, the aim of the present study was to investigate the DNA methylation level of AR and its coregulators, especially those mapped on the X-chromosome, that may influence the activity of AR in MBC.Methods: The DNA methylation level of AR, MAGEA2, MAGEA11, MAGEC1, MAGEC2, FLNA, HDAC6, and UXT, mapped on the X-chromosome, was evaluated by quantitative bisulfite-NGS. Bioinformatic analysis was performed in a Galaxy Project environment using BWA-METH, MethylDackel, and Methylation Plotter tools. The study population consisted of MBC (41 cases) compared with gynecomastia (17 cases).Results:MAGEA family members, especially MAGEA2, MAGEA11, MAGEC, and UXT and HDAC6 showed hypomethylation of several CpGs, reaching statistical significance by the Kruskal–Wallis test (p < 0.01) in MBC when compared to gynecomastia. AR showed almost no methylation at all.Conclusions: Our study demonstrated for the first time that MAGEA family members mapped on the X-chromosome and coregulators of AR are hypomethylated in MBC. This may lead to their overexpression, enhancing AR activity. Even if its incidence is slightly increasing, male breast cancer (MBC) is a rare disease accounting for about 1% of all breast cancers –4 and beRecent papers based on molecular analyses of large multi-institutional series demonstrated that, in spite of some similarities with FBC, MBC shows a specific molecular portrait , 5–7.AR) gene is emerging as playing a key role in male breast neoplastic transformation  family genes, all mapping to the X-chromosome  in MBC. In addition, AR regulator genes on the X-chromosome like FLNA, HDAC6, and UXT were studied. Results obtained in invasive MBC were compared with gynecomastia as controls.The aim of this study was therefore to evaluate the methylation level of N = 17) were selected when not associated with invasive carcinoma, either synchronous or metachronous.MBC and gynecomastia cases were retrieved from the files of the Pathology Units of the Universities of Bologna , Rome , Italy, Zurich , Switzerland, and Utrecht, The Netherlands. Tissues had been routinely formalin-fixed and paraffin-embedded (FFPE). Cases were retained when enough informative DNA was obtained from the FFPE tissue samples. Gynecomastia cases  applying a monoclonal antibody .All clinical investigations have been conducted according to the principles expressed in the Declaration of Helsinki. The study was approved by local Ethics Committee of Bologna (protocol number CE-AVEC 17180). Further use of cases was approved by the local ethical committees of Zurich (KEK_2012-553 and KEK-2012-554) and Utrecht . After a denaturation step at 95°C for 5 min, the solution was centrifuged at 10,000 × g at 4°C for 5 min. The interphase containing DNA was quantified by Nanodrop  and stored at 4°C or immediately processed for the bisulfite-NGS protocol.DNA purification was performed as previously described  and summAR, MAGEA2, MAGEA11, MAGEC1, MAGEC2, FLNA, HDAC6, UXT, all mapped on the X-chromosome. Locus-specific bisulfite amplicon libraries were generated with tagged primers using Phusion U DNA polymerase  and loaded onto MiSEQ  according to the manufacturer's protocol. FASTQ output files were processed for quality control  and converted into FASTA format in a Galaxy Project environment (http://cbc.case.edu/BSPAT/index.jsp) (U-test using the Methylation Plotter tool available online (Bisulfite treatment of genomic DNA (100–500 ng) was carried out with the EZDNA Methylation-Lightning™ Kit  according to the manufacturer's protocol. Quantitative DNA methylation analysis was performed as previously described  using a dex.jsp) , Kismethdex.jsp) , and fine online . Principe online .The study population comprised 41 men with a mean age of 63 years (range 49–93). All cases were diagnosed as invasive carcinoma, no special type (IC-NST), and were ER and PR positive. No HER2 amplified cases were included. Clinic-pathological details are reported in AR was expressed in all cases. AR antibody stained 10–100% of the neoplastic cells  in MBC compared to gynecomastia  and the second highest principal component , cases are distributed considering the methylation level of the total of 92 CpGs .The HeatMap  generatein situ carcinoma as well as in cancer-associated gynecomastia  regulates the cytoskeleton organization by linking with actin filaments. In addition, FLNA protein interacting with >60 different other proteins can regulate several cell functions ranging from cell migration, transmembrane receptor signaling to DNA damage repair , 25. AmoFLNA in MBC. According to the present results, the FLNA gene extracopies are all hypomethylated and therefore likely transcriptionally active.No data have been previously published on ER transcriptional activity  mapped to Xp11.23, also called androgen receptor trapped clone-27 , is a prHDAC6) mapped to Xp11.23 is an important AR regulator in prostatic cancer by enhancing AR protein stability  (https://www.ebi.ac.uk/ena).The original contributions presented in the study are publicly available. This data can be found here: European Nucleotide Archive . Further use of cases were approved by the local ethical committees of Zurich (KEK_2012-553 and KEK-2012-554). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.LMor, RM, and MF contributed to the conception and the design of the study. GG organized the database. LMor and MF wrote the first draft of the manuscript. GG, CM, and LMor performed molecular analyses of the tissues. AMS, MC, and AM contributed to case recruitment and data discussion. AS, GZ, ZV, LMos, and PD revised the cases and contributed to the case selection. All authors contributed to manuscript revision, and read and approved the submitted version.MF received funds from Roche and Devicor Mammotome, for course organization and from Biocartis and MSD for professional consultation. None of the funds recevied are related to the present study or influenced the results. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Renal cell carcinoma (RCC) is one of the most common malignant tumors originating from the renal parenchymal urinary epithelial system. Tripartite motif 47 (TRIM47) is a member of the TRIM family proteins, which has E3 ligase activity and has been demonstrated to be involved in the occurrence and prognosis of many tumors. The main purpose of this study is to explore the role and potential mechanism of TRIM47 in promoting malignant biological behavior of RCC.TRIM47 mRNA and protein levels in human renal cancer and paired normal adjacent tissues were detected by qRT-PCR and Western blot. The effects of TRIM47 knockdown and overexpression in renal cell carcinoma cells on cell proliferation, invasion and xenograft tumor growth in nude mice were analyzed. The molecular mechanism was explored by mass spectrometric exploration,Western blot and immunoprecipitation assays.TRIM47 promoted RCC cell proliferation in vitro and in vivo as an oncogene. Mechanistically, TRIM47 exerted an E3 ligase activity by interacting with P53 protein to increase its ubiquitination and degradation, which further promoted the malignant biological behavior of RCC.Our study demonstrated that the TRIM47-P53 axis played a functional role in RCC progression and suggested a potential therapeutic target for RCC. Renal cell carcinoma (RCC) is a common malignant tumor of the urogenital system, the incidence of which is only next to that of prostate cancer and bladder cancer, accounting for 2.2% of adult malignant tumors . At presThe advent of targeted drugs as adjuvant therapy and their promising therapeutic efficacy have brought new hope for patients with high-risk and locally advanced RCC patients. Tyrosine kinase inhibitors (TKIs), such as sunitinib, axitinib, perzopanib and sorafenib, are the most commonly used molecular targeted drugs for kidney cancer. They are vascular endothelial growth factor receptors (VEGFRs) that reduce tumor cell growth by targeting the tumor angiogenesis pathway , 5. HoweTripartite motif-47 (TRIM47) is a member of the TRIM protein family. The most important structure in this motif is a really interesting new gene (RING) domain. Proteins with a RING domain often play the role of E3 ligase activity –10. TRIMThe purpose of this study was to further determine the role of TRIM47 in the development and progression of RCC by immunohistochemical staining of RCC specimens, construction of TRIM47-overexpression plasmids, lentiviral shTRIM47 knockdown cell line, and Crispr/cas9 technology to construct knockout methods such as TRIM47 stably transformed strains, animal experiments, mass spectrometry detection, bimolecular fluorescent complimentary experiments and co-immunoprecipitation experiments, in an attempt to gain deeper insights into the molecular biological function and mechanism of TRIM47 in RCC.Clinical specimens of kidney cancer and paired adjacent tissues were collected from 6 patients who received surgical treatment between October 2017 and February 2018 at the Department of Urology of the Oriental Hepatobiliary Surgery Hospital affiliated to the Naval Medical University . They included 4 pairs of clear cell renal cell carcinoma (CCRCC), one pair of papillary renal cell carcinoma (PRCC) and one pair of chromophobe renal cell carcinoma (CRCC). The postoperative clinical specimens of 125 patients constituted paired RCC and paired normal adjacent tissues. Clinicopathological variables including age at age, gender, tumor size, pathological types and tumor stages were analyzed  and normal renal cell HK-2  were routinely maintained in RPMI-1640 , DMEM (Gibco) and McCoy’s 5A supplemented with 10% FBS (Gibco), 100 U/mL penicillin sodium and 100 mg/mL streptomycin in a humid atmosphere with 5% COAfter dewaxing, hydration and antigen repair, the renal cell carcinoma tissue chip was added dropwise with diluted rabbit anti-TRIM47 antibody  and incubated overnight. The diluted anti-rabbit HRP and horseradish enzyme-labeled streptavidin working solution were added dropwise to the tissue chip the next day. The tissue chip was then stained with DAB chromogen and counterstained with hematoxylin, dehydrated and observed and counted under an optical microscope. The results of histochemical staining and the evaluation criteria of the staining intensity were scored as 0 for negative, 1 for weakly positive, 2 for moderately positive, and 3 for strongly positive. The evaluation criteria for positive cell frequency were as follows: < 5%, 0 point; 5–25%, 1 point; 26–50%, 2 points; 51–75%, 3 points; and > 75%, 4 points.The coding sequence of TRIM47 was synthesized by TSINGKE Biological Technology , and then inserted into pcDNA3.1(+) using the Quick-Fusion cloning kit .The shRNA sequence of TRIM47 was CAAGAAGTCCTGCATATCCGT. After 293T cells were infected with the lentivirus packaging plasmid, and 769-P and A498 cells were infected with the virus for 48 h, they were observed under a fluorescence microscope.TRIZOL  was used to extract the total RNA, and then reverse transcribed into cDNA. Finally, SYBR Premix Ex TaqTM II  was used to quantify gene transcripts on the 7900HT fast RT-PCR system  and standardized with GAPDH.Primer sequence: GAPDH-F: GTCTCCTCTGACTTCAACAGCG; GAPDH-R: ACCACCCTGTTGCTGTAGCCAA; TRIM47-qF: ACGGCAGTGGACCCTTCAG; TRIM47-qR: CCAGGCAGGCGAGACAGAA.Total protein was first extracted using SDS-PAGE and then transferred to a PVDF membrane . Afterwards, the PVDF membrane was incubated with 1:1000 dilution of antibody: TRIM47 , BAP1 , P21 , P53 , SRC , MET , and c-Myc . The membrane was washed and incubated with a 1:2000 dilution of horseradish peroxidase-conjugated goat anti-rabbit . SuperSignalMT West Puico PLUS  was used to develop the blot, using b-actin  as the loading control. All experiments were performed in triplicate.After digestion, counting and centrifugation, overexpression cells or knockdown cells were seeded into 96-well plates with 5000 cells per well, cultured for 24 h, and finally evaluated using the Cell Counting Kit 8 . The absorbance at 450 nm was measured using an ELx800 plate reader .5/mL. The bottom of the chamber is 1640 medium containing 15% FBS, which is placed at  for 48 h. Cells were fixed in 10% methanol solution for 30 min, stained with 0.1% crystal violet for 20 min, and washed in tap water until the background was clear. Five fields were randomly selected under the light microscope, and the number of membrane-passing cells was counted.Matrigel was placed on ice in advance to hydrate overnight, and then diluted with serum-free medium at a 1:8 ratio, followed by addition of 50 µl matrigel to the bottom of the transwell chamber. Cells were digested, counted and then inoculated onto the transwell chamber. The total amount of cells per well was 0.4 × 10Crispr/Cas9 system was used to construct the knockout-TRIM47-769P cell. After constructing the sgRNA-cas9 expression vector and transfecting the 769P cell line, the stable transgenic strain was constructed through the lentiviral packaging system and selected by puromycin. The sgRNA sequence of TRIM47 is CGCCTGCCTGGGCGCGCTCTGG.7/mL, each mouse was injected with 0.2 mL cells (about 2 million cells) into the upper right thigh and observed for 3 weeks.The experimental procedures were approved by the Second Military Medical University. Male BALB/c-nu mice  were purchased from Shanghai Laboratory Animal Company . Sixteen nude mice were equally randomized into two groups: WT-769P group and KO-TRIM47-769P group. After adjustment of the concentration to 1 × 10The TRIM47 sequence was inserted into the Flag-tagged vector, and the P53 sequence was inserted into the His-tagged vector to form two recombinant plasmids. The recombinant plasmid carrying the target gene and the virus packaging plasmid were co-transfected into 293T cells and then infected 769-P cells. The corresponding stable transfected cells were screened using puromycin. The His-P53 overexpression stable transfected cells and His-P53 + Flag-TRIM47 overexpression stable transfected cells were obtained. After the extraction of the immunoprecipitated protein, the antibody was combined with protein A/G agarose, and then the bound antibody was crosslinked, followed by pre-clearing of the protein sample, antigen immunoprecipitation and antigen elution before Western Blot detection was performed.Statistical data are expressed as means ± standard deviation (SD). All data were statistically analyzed and processed using SPSS (IBM SPSS Statistics 22.0) software. The experiment was repeated three times and then analyzed statistically.The results of Western Blot detection of the fresh RCC and matched normal adjacent tissues indicated that the expression level of TRIM47 protein in RCC tissues was significantly higher than that in adjacent tissues Fig. a. IHC stTo clarify the biological effect of TRIM47 in RCC, we firstly detected the expression of TRIM47 in 6 different RCC cell lines and normal renal cell HK2 by using Western blot and qRT-PCR, and the results showed that 769P and A498 cells highly expressed TRIM47 while 786-O and ACHN cells lowly expressed TRIM47 Fig. a, b -769P group as control. After 3 weeks of the tumor-bearing experiment in mice, the tumor proliferation ability in KO-TRIM47-769P group was reduced significantly as compared with that in WT-769P group (Fig. Members of the TRIM protein family are known to be involved in many biological processes, and changes in abundance or activity are related to a variety of pathological conditions, including viral infections, developmental and neurodegenerative diseases, and cancer occurrence . Recent IHC analysis of the RCC and matched adjacent tissue chips showed that the expression of TRIM47 was up-regulated in RCC tissues. Analysis of TCGA data showed that the expression level of TRIM47 was related to the clinical prognosis of RCC patients, and OS of the patients with high expression was lower than that in the patients with low expression. After overexpression of TRIM47, the proliferation and invasion abilities of 786-O and ACHN RCC cells were significantly enhanced. On the contrary, TRIM47 knockdown mediated by lentiviral shRNA decreased the proliferation and invasion abilities of 769-P and A498 RCC cells significantly. The results of the animal model showed that the use of Crispr/cas9 technology to knock out TRIM47 reduced the tumor-bearing ability of nude mice. The above evidence shows that TRIM47 can promote the malignant formation of RCC, suggesting that it may prove to be a potential biomarker for RCC. We then used MS to compare WT-769P with KO-TRIM47-769P. The results showed that there were many significantly differentially expressed proteins between the two types of kidney cancer cell lines, based on which, we selected 6 protein molecules with significant differences, including P53, P21, BAP1 common tumor suppressor proteins, SRC, MET and c-Myc common oncoproteins. Subsequent Western Blot assay showed showed that the protein levels of P53, P21, and BAP1 were significantly down-regulated in RCC 769-P cells after TRIM47 overexpression, and vise versa after TRIM47 knockout. At the same time, the protein levels of SRC, MET and c-Myc were increased after TRIM47 overexpression, and increased after TRIM47 knockout.The p53 gene is the most common human tumor suppressor gene. It is located on the human chromosome 17p13.1. As its protein band is located at 53KDa, it is named P53 protein or TP53 protein. P53 protein has the ability to induce cell cycle arrest, senescence and / or apoptosis, which is the core of its anti-tumor mechanism , 26. TheIn summary, TRIM47 plays a post-translational modification role in RCC by exerting an E3 ligase activity, mediating ubiquitination and degradation of P53 tumor suppressor protein and promoting malignant progression of RCC cells at the protein level. The role of TRIM47 as a tumor-promoting molecule in promoting the malignant biological behavior of RCC is crucial. However, the specific action mechanism of TRIM47 in RCC needs further exploration. TRIM47 overexpression may prove to be a useful prognostic factor and a potential treatment target for RCC.


Stroke is a disease affecting a large part of our society. According to WHO data, it is the second world’s biggest killer, accounting for near six million deaths in 2016 and it is about 30% of the total number of strokes per year. Other patients affected by such a disease should be rehabilitated as soon as possible. As a result of this phenomenon, paresis may occur. Among the devices available on the market there are many rehabilitation robots, but the method of electrostimulation can be used. The authors focused their attention on electrostimulation and commercially available therapies. Using this method, application to people with large hand muscle contracture is difficult. The authors of the work present a solution dedicated to exactly such people. A solution of textronic sensors manufactured on a textile substrate using the technology of physical vapor deposition is presented in the article. As a result of the conducted research, an electroconductive structure was obtained with a low surface resistance value of 1 Ω/□ and high flexibility. It can alternatively be used in hand rehabilitation for electrostimulation of fingertips. The solution is dedicated to people with high hands spasticity for whom it is impossible to put on a rehabilitation glove. Stroke is a neurological disease that affects mainly elderly people. Currently, the number of people suffering from the first strokes has increased approximately by 68% in Poland in the period time 1990–2016 and the number of disabled people is also increasing largely in the world.The paresis of the upper limbs may be the result of ischemic stroke, which is the most common cause of permanent inability to work and perform basic life functions. Ischemic strokes account for over 80% of all strokes, while others are hemorrhagic strokes .According to Saver , the celMovement dysfunction is one of the key problems after stroke. Motor relearning during rehabilitation programs is based on repetition. Therefore, intensive and repetitive task-specific training, which involves the active practice of task-specific motor activities, is recommended after stroke. There are many different factors that have an impact on neuroplasticity after stroke. One of the most important factors on which the effectiveness of the therapy depends is the early implementation of treatment and rehabilitation. Individually-tailored rehabilitation to each patient with different abilities, functional needs and interests is the most important challenge in post-stroke care. The key time is limited to a critical period of six months. Due to the high plasticity of the brain, immediate and long-term rehabilitation allows reducing the neurological deficit. The restoration of brain functions is based on the distributed functional neural networks, which are located in the regions that are not touched by the brain infarction.The next factors are regularity, repeatability, intensity of performed exercises. In the literature, we can find several methods that, when used together, bring the best results. Repetitive task training (RTT), Constraint-Induced Movement Therapy (CIMT), Strength Training, Aerobic Training, Non-Invasive Brain Stimulation (NIBS), Computer-Based Virtual Reality Training or Neuromuscular Electrical Stimulation.Currently, we can also notice some major developments and innovations of Robotic-Assisted Rehabilitation (RR) and Robotic-Assisted Therapy (RT). From year to year, we observe an increase in the number of next-generation rehabilitation robots that can lead to significant benefits in neurorehabilitation. Using RT, we can measure several parameters that are usually not considered in conventional therapy. They include inter alia, spasticity, reflexes, level of voluntary control and functional movements. In addition, the therapy guarantees high intensity of repetitive movement training, task-specific and interactive treatment and monitoring a patient’s progress. Therefore, it is a perfect tool for doctors and therapists for facing neurorehabilitation challenges. Furthermore, robot-assisted therapy can also significantly decrease the labor intensity and costs associated with recovery training .The rehabilitation assisted by robots and mechanical devices, because of their intended use, can be divided into two groups supporting the lower- and upper-limb rehabilitation. Some of them like the Robot RRH1, created in the Institute of Automatic Control Technical University of Lodz, may be used even for people lying in beds . ResearcThe second group is the therapy of the paresis of the upper limb. That one is considered the most difficult part of the process of rehabilitating patients after a stroke. We can divide the problems into two groups. One is a decrease in muscle tone (plegic) usually appearing immediately after a stroke, the other is a permanent limb cramp (spasticity) which is a consequence of giving up hand exercises when there is a decrease in muscle tension and it prevents any precise movement of the hand. .Although many robotic rehabilitation devices have been developed and introduced at the market, e.g., , there iAnother common problem is hand rehabilitation. High finger mobility allows us to grab, pinch, manipulate and interact with objects. Good, efficient hands without paresis are the basis for normal functioning in the environment in daily life. Additionally, in that case, to prevent permanent spasticity, post-stroke therapy should be carried out immediately after stroke. In the literature, we can find descriptions of many devices created for hand therapy of people with paresis ,11,12. OThe second group is based on a small size and portability solution of the system, which could allow the users to undergo rehabilitation on their own in their homes. Designed small robotic manipulators, so-called intelligent glows, mechanically supported tools to straighten and bend the hand, can decrease physiotherapist’s involvement during patients’ demanding treatment. Using tools like that can result in increasing the treatment quality and the number of patients rehabilitated at the same time. These popular devices are intended for people with neurological and orthopedic injuries resulting from stroke, Parkinson’s disease, musculoskeletal disorders and upper limb injuries. The system relies on the neuroplastic abilities of nerve tissue to create new connections and self-repair by stimulating the proprioceptive system. We can find many examples of mechatronic systems like the “Wearable hand rehabilitation system with force feedback” project being developed at the Institute of Automation and Robotics, Poznan University of Technology , based oIt is very important for stroke patients to improve the quality of hand function. In many cases, patients after stroke have post-stroke spasticity. Upper-limb rehabilitation is not sufficient to restore hand function. There is a solution on the market in the form of a textronic glove developed by scientists from the Institute of Neuroinformatics  of the University of Bochum, Germany. According to their solution, a patient with hand paresis can be rehabilitated using a TipStim© glove. However, this solution is not satisfactory for people with high spasticity, for whom it is impossible to put on a glove .The authors of the work propose an innovative solution consisting in the production of electric sensors on a composite elastic textile substrate, which can be used in an existing system of textronic gloves.Different types of diagnostic sensors can be integrated into textiles with such methods such as: embroidery, physical vapor deposition, sputtering, chemical vacuum deposition, sol–gel, inkjet printing or polymer-assisted physical vapor deposition ,17,18. IThe percentage of e-textile players that use various technologies in the production of textronic solutions is shown in Wearable electronics solutions include sensors using changes in electrical parameters, such as impedance , inductaMany of the wearable sensors are in direct contact with human skin. For this reason, they should provide wearing comfort as for example the ACC/PAA/alginate hydrogel-based ionic skin sensor developed by Lei et al. which can be applied to detect gentle finger touch, complicated muscle movements during speaking, detect human motion or blood pressure . The preSuch systems can be used in various medical tests, such as electrocardiography (ECG), electromyography (EMG) and electroencephalography (EEG), pH level, temperature, blood pressure, breathing and many others. The multi-perspective view on patient data is necessary to improve medical decision making . All theThe new approach in electric stimulation using a wearable system has been mastered very recently. It is the electric stimulation of tips. The treatment is realized using two-directional pulse currents with a long interval time between pulses. It is assumed that the stimulation of tips evokes some change in neuronal cells in the patient brain. Technically, the stimulation is realized by a five-finger glove with electrodes installed on the tips . The glove is equipped with two independent electric circuits. One of them is used to stimulate the median nerve. In this case, the electrodes are placed on the distal and middle phalanges of fingers 1–3. The second electrical circuit applies to 4–5 fingers and is used to stimulate the ulnar nerve . The valAfter long-term research and searching for a material that could act as a base for the stimulation electrode, an Optisana© bandage was chosen. It is produced on the basis of transparent nonwoven fabric which is made of loosely braided polyurethane threads. A thin electroconductive layer was produced on that substrate in the process of physical vapor deposition. The parameters of the metallic layer allow for implementing it into the TipStim glove systemAll microscopic images were collected with the optical KEYENCE VK-X1000 microscope. The magnification of the tested structures is marked in the pictures.−5 mbar initial vacuum and lasted 5 min. Silver with a high conductivity value (γ = 62.5 m/(Ω·mm2)) and high purity provided by Polish Mint Ltd. was used as a deposited material. The vacuum deposition process was carried out using a resistive resistance source made of tungsten, characterized by a much higher melting temperature than silver. The distance between the evaporation source and the surface on which the metallic conductive layer was deposited was 6 cm. Increasing this distance resulted in the inability to obtain a continuous layer with a low resistance value. Reducing the distance resulted in the patch overheating. Prior to the technological process, fibrous substrates were pre-conditioned for several hours at room temperature and 55% humidity. The connecting wires were attached to the sensor structure using ELPOX AX 15S electrically conductive glue  with low resistivity (1.6 × 10−6 Ωm). Contact modification can be performed using laser ablation of its surface [A thin electroconductive layer was created in the process of physical vapor deposition using the Classic 250 vacuum chamber of the Pfeiffer Vacuum system. The process started after reaching 5 × 10 surface ,23,24.In the first stage of research, the surface structure of the electrodes used in the existing solution for rehabilitation was analyzed, as well as the structure of fibrous material present in the bandage, and used as a substrate to produce the median and ulnar nerves’ electrostimulation electrode. An image of the surface of the material used in the rehabilitation glove as an electrode is shown in Despite the lack of an organized weave between the threads in the chosen medical patch, a continuous metallic layer with a surface resistance of 1 Ω/□ was created as a result of physical vapor deposition. This resistance was measured in a typical four-electrode system .The structure and surface profile of the embedded surface are shown in Rmax) is 332.74 µm [The analysis of the microscopic image shows that, despite significant unevenness on the surface of the fibrous substrate, a thin-film structure with electrocontinuity was created. Atoms of stochastically deposited metal penetrated deep into the material creating a thin metallic layer. The roughness profile shown in 32.74 µm . Using pA 36-year-old patient was admitted to rehabilitation and was admitted to the Rheumatology Clinic due to persistent paresis of the left upper limb, slight paresis of the lower limb. The patient was confirmed ischemic stroke of the right hemisphere during MRI magnetic resonance imaging. After the implementation of treatment and stabilization of the patient at the Rheumatology Clinic of the Military Medical Institute, she was redirected to rehabilitation treatment at the Rehabilitation Clinic. Then, after diagnosing deep sensation disorders while having preserved superficial sensation, she was qualified for electrostimulation using a TipStim glove.Rehabilitation was carried out using the PNF (Proprioceptive Neuromuscular Facilitation) method for 120 min six times a week and, additionally, the fingertips were electrostimulated using a TipStim glove for 60 min with a frequency of once a day excluding Sundays for four weeks. After this period, the patient was again subjected to dexterity tests. The applied functional hand tests were aimed at an objective assessment of small and large motor functions of the hands using simulated everyday activities as well as muscle strength and the return of superficial and deep sensation. In addition, a comparison was made of the outcome of the patient’s therapy after a stroke resulting from systemic vasculitis with a healthy person of a similar age and gender. After four weeks of rehabilitation of a stroke patient the use of fingertip electrostimulation, significant motor limb improvement with paresis was obtained . It confAmong the many technical solutions supporting the rehabilitation of people after stroke who have spasticity of the hands, solutions dedicated to supporting the restoration of the motor functions of the hands and palms, should be noted. In some patients, the level of paralysis of the nerves of the hand is so great that it is impossible to take advantage of the solutions offered by robotics. In this case, the solution adequate to such applications is rehabilitation using the TipStim glove with newly developed electrodes for electrostimulation. Thanks to the flexible substrate used as well as the process of physical vapor deposition, the produced electrodes are flexible and allow sensors to be worn by a person with any degree of dexterity of the hand.In order to use electroconductive threads available on the market to create electrodes for finger electrostimulation, such a solution should be designed at the initial stage of glove design. Creating electrodes for electrostimulation of fingers based on dressing patches will allow personalizing the electrodes and adapt them to the needs of the patient. The electrodes produced in the PVD process will be irreplaceable in the situation of adjustment to the individual needs of the patient taking into account the size of the fingers. Using the proposed technology for producing a thin electroconductive layer on the proposed substrate, it is also possible to produce electrode matrices of the desired dimensions in one technological process by masking or laser ablation of the metal from the resulting structure.In addition, it should be emphasized that no continuous electrical conductivity of the produced structures on textile substrates characterized by spatial structure is observed. In the described case, in the technological process of physical vapor deposition, obtaining thin electroconductive layers on such substrates was successful.The authors of this study suggest using the method of electrostimulation of fingertips using the TipStim glove with the proposed new electrodes in the rehabilitation of patients with paresis of the upper limb after a stroke in order to improve superficial and deep sensation and improve the efficiency of the hand. The legitimacy of its use was confirmed by the therapies carried out for people with paresis.


Patients diagnosed as having AR exhibited higher comorbidity rates, including of asthma, COPD, CRS, gastroesophageal reflux disease, and nasal septum deviation, than those of the comparison cohort. Our results strongly indicate that AR is an independent risk factor for laryngeal pathology. Therefore, when treating AR and voice problems, physicians should be attuned to possible laryngeal pathology.Allergic rhinitis (AR) is correlated with diseases including allergic laryngitis, chronic obstructive pulmonary disease (COPD), asthma, and chronic rhinosinusitis (CRS). The unified airway model suggests that inflammation can spread in both lower and upper respiratory tracts. Moreover, some voice problems—laryngeal edema, dysphonia, and vocal nodules—have been associated with AR. We examined the association between AR and laryngeal pathology. We investigated 51,618 patients with AR between 1 January 1997 and 31 December 2013, along with 206,472 patients without AR matched based on age, gender, urbanization level, and socioeconomic status at a 1:4 ratio. We followed patients up to the end of 2013 or their death. The occurrence of laryngeal pathology was the primary outcome. Individuals with AR had a 2.43 times higher risk of laryngeal pathology than the comparison cohort group (adjusted HR: 2.43, 95% CI: 2.36–2.50,  Allergic rhinitis (AR), which affects nearly 40% of people worldwide, is a growing public health concern. AR is commonly associated with asthma  under thVoice problems affect up to 41 million older adults in the United States, and among younger adults, a prevalence of 6% has been noted . VocalizAccording to the literature, fiber optic endoscopic examination has revealed that patients with AR are affected by voice symptoms and chronic edema of the vocal fold ,9. MoreoWe adopted a nationwide cohort study design, employing population-based data obtained from the NHIRD. In 1995, the Taiwanese government established its NHI program, which is a universal scheme covering medical care for more than 99% of the residents of Taiwan . BecauseApproval for this study was granted by the Institutional Review Board of Chang Gung Medical Foundation . NHIRD data do not contain personally identifiable information, and thus, privacy of the enrolled sample was ensured. The NHI Administration and the Chang Gung Medical Foundation, through its Institutional Review Board, guaranteed that all heath and personal information would remain confidential.Our analyses were conducted on 51,618 patients with AR and 206,472 comparison patients without AR; these cohorts were matched by sex, age, socioeconomic status, and urbanization level at a 1:4 ratio. The procedure for selecting participants is illustrated in For enrollment in the comparison group, we randomly selected patients without AR from the LHID2000. According to age, urbanization level, sex, and socioeconomic status, we matched these comparison patients with AR patients at a 1:4 ratio.We followed patients until they died or the study period ended (31 December 2013). Laryngeal pathology was the primary outcome of interest .The sociodemographic information of patients  was retrieved from the initial enrollment data. We searched the outpatient and inpatient claims data for comorbidities related to AR, specifically asthma   for laryngeal pathology. Subgroup analyses were conducted to identify potential effect modifiers and were stratified by sex, age, and comorbidities. SAS v. 9.4  was applied for all statistical analyses, and a two-tailed Our results indicated that individuals in the AR cohort had a higher likelihood of having laryngeal pathology and the comorbidities of asthma, COPD, CRS, and GERD than patients in the comparison cohort . Moreovep < 0.001; We used the Kaplan–Meier approach to identify a cumulative incidence of laryngeal pathology that was higher at the significance level in the AR cohort compared with the cohort without AR , we were able to identify the trends and changes in laryngeal pathology risk in both cohorts. Moreover, patients who had received a diagnosis of laryngeal pathology prior to their AR diagnosis were excluded, which helped to clarify the sequence of disease occurrence. We compared the results of the study and comparison cohorts by using matching Cox models after adjustment for comorbidities; thus, we were able to account for the influence of potential confounders. In addition, the effect of AR was consistent after subgroup analyses; we determined that, in all subgroups, laryngeal pathology risk was higher in patients with AR than in those without. Therefore, we confirmed AR to be an independent risk factor of laryngeal pathology.AR is a commonly encountered inflammatory disease affecting the nasal mucosa. Some studies have found that AR is involved in systemic inflammation and is closely related to other inflammatory diseases and conditions, including rhinosinusitis, asthma, and allergic conjunctivitis . FurtherThe laryngeal pathology of individuals with AR or laryngitis has only been insufficiently investigated, possibly because laryngoscopy is not a routine procedure when diagnosing AR or laryngitis. AR causes nasal congestion, nasal obstruction, and mucus hypersecretion, and triggers rhinolaryngeal reflexes, resulting in altered voice resonance, hoarseness, laryngeal edema, and dysphagia . In one We demonstrated that, compared with the non-AR cohort, the incidence rate of laryngeal pathology was higher at the significance level in the AR group. Moreover, after adjustment of some covariates, such as asthma, COPD, CRS, GERD, and NSD, the multivariate analysis revealed that AR group showed statistical significance in all subgroups. Some studies have determined that patients with AR frequently experience the comorbidities of asthma, COPD, CRS, and GERD; our results are in accordance with these findings ,19. WhenSeveral limitations should be mentioned. First, this was a retrospective study in which we used population-based data; we lacked the original medical records to confirm diagnoses. Therefore, we used the criteria of at least three outpatient diagnoses or one inpatient diagnosis to strictly define the diseases. Second, patient symptoms, medication, smoking history, laboratory data, including serum immunoglobulin E and G, and specific allergen testing data were not available in this database; thus, we could not determine the severity of AR. Third, the diagnosis of laryngeal pathologies is based on ICD-9-CM codes, but the laryngoscopy imaging and pathology report were not available in the database; consequently, we could not diagnose laryngeal pathology in more detail. Finally, the definite mechanism responsible for the association between AR and laryngeal pathology requires further study.We investigated the relationship between laryngeal pathology and AR using a national population-based database and determined AR to be an independent risk factor of laryngeal pathology. Consequently, we suggest that clinicians should be attuned to the possibility of laryngeal pathology in patients with AR. Moreover, when treating patients with AR and voice problems, a thorough laryngoscopy is suggested.


Our goal was to evaluate the efficacy of muscle spasm before and after rehabilitation by comparing shear wave propagation velocity (SWV) and Young's modulus (YM) in the normal and spastic biceps brachii in stroke patients.  A study of 60 stroke patients with upper limb spasm was performed; these patients were admitted from April 2018 to September 2019. The modified Ashworth scale (MAS) scores of the spastic biceps brachii before and after rehabilitation treatment were compared. SWV and YM on the spastic and normal biceps brachii before rehabilitation treatment, SWV and YM on the spastic and normal biceps brachii after rehabilitation treatment, and SWV and YM on the spastic biceps brachii before and after rehabilitation treatment were compared. Whether SWV and YM on the spastic biceps brachii are related to MAS was compared. P < 0.01) and after (P < 0.05) the rehabilitation treatment. There was no statistically significant difference in SWV and YM in the normal biceps brachii before and after the rehabilitation treatment (P > 0.05). There was a statistically significant difference in SWV and YM in the spastic biceps brachii before and after the rehabilitation treatment (P < 0.01). SWV and FM of the spastic biceps brachii are correlated with MAS before and after rehabilitation treatment, and the correlation coefficient for SWV was 0.563 and 0.605 for YM (P < 0.05).  There was a statistically significant difference in SWV and YM between the normal and spastic biceps brachii before ( SWE can be used as a means of assessment before and after rehabilitation treatment. Stroke is characterized by high incidence, and the patients have high disability rates. A large number of studies have shown that the incidence of upper limb spasm is significantly higher than that of the lower limb, with a more difficult rehabilitation –2. SpastAt present, the assessment of dysfunction in patients with upper limb spastic paralysis is mostly based on subjective scale assessment, which is inaccurate and not sufficiently sensitive . Muscle All study procedures involving human participants were in accordance with the ethical standards of the institutional and national research committee and the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. This study was approved by the hospital institutional review board. Oral informed consent was obtained from all individual participants included in the study, which was approved by the hospital institutional review board. Sixty stroke patients with hemiplegia and one-sided upper limb spasms were selected between April 2018 and September 2019. Participants included 36 males and 24 females. Patient ages ranged from 56 to 78 years, with an average of 66 ± 7.46 years. The course of stroke ranged from 1 to 6 months, with an average of 6.6 months. There were 48 cases of cerebral infarction, 12 cases of brain bleeding, 35 cases of left hemiplegia, and 25 cases of right hemiplegia. Patients with no upper limb spasm served as the control group.The inclusion criteria were as follows: (1) patients with unilateral hemiplegia after the first stroke between 1 and 6 months, (2) cerebral hemorrhage or cerebral infarction confirmed by computer tomography or magnetic resonance imaging, (3) upper limb spasm on one side and no previous myogenic disease, (4) patients with elevated upper muscle tone and upper limbs who were able to remain in a stretched position and the modified Ashworth scale (MAS) graded 0–3, and (5) patients who can cooperate with the examination and assessment.The exclusion criteria were as follows: (1) dystonia, such as Parkinson's syndrome; (2) paralysis of bilateral limbs, such as increased muscle tone due to spinal cord injury; (3) complications of serious medical illness or mental or cognitive impairment, such that the patient cannot cooperate with examination; and (4) MAS graded 4 and patient with stiff muscles who was unable to stretch and flex.MAS scores were as follows: level 0—0 points, level 1—1 point, level 1+—2 points, level 2—3 points, level 3—4 points, and level 4—5 points. The total MAS scores in the spastic side before and after the rehabilitation treatment were compared.This study was performed using a sonography scanner incorporating a 9L probe (LOGIQ E9) with a frequency of 9 MHz.All patients were placed in a supine position, with the upper limbs of the spastic side positioned at 90 degrees relative to the body and biceps brachii in a stretched position. First, the probe was used to find the largest cross-section of the musculoabdomen biceps, and then it was rotated 90 degrees, parallel to the direction of the muscle bundle. SWE was then performed within an ~10 × 10 mm area of interest and a depth of 1–3 cm. After the elastic image was stabilized, the Q-BOX function was frozen to measure the SWV (m/s) and YM (kPa) of the muscle tissue in the region of interest. The measuring circle diameter was 2 mm, and the same location was measured three times to obtain an arithmetical average value. After the spastic side test was completed, the normal side test was performed according to the above method. The above examination after the rehabilitation treatment was repeated, and the bilateral SWV and YM were compared. The SWV and YM correlation was analyzed using MAS.Routine rehabilitation treatment utilized Bobath technology and relearning exercises once a day using one-on-one training with a professional rehabilitation therapist 45 min each time. The remaining training was practiced by the patients themselves. Functional electrical stimulation used two electrode plates placed on the triceps brachii to trigger elbow extension . One electrode plate was placed on the back of the deltoid muscle and the other was placed on the supraspinatus muscle to trigger shoulder lift and abduction based on patient tolerance. The treatment was performed for 30 min each time, two times a day for a total treatment time of six weeks.P < 0.05 was considered statistically significant.The data were expressed as the mean value ± one standard deviation using SPSS19.0 software. An independent sample nonparametric test was used to compare between and within groups. Spearman's test was used for the correlation analysis of SWV and YM with MAS. P < 0.05)  .P < 0.01) and after (P < 0.05) (P > 0.05). There was a statistically significant difference in SWV and YM in spastic biceps brachii before and after the rehabilitation treatment (P < 0.01)   the reha < 0.01) .P < 0.05). The correlation coefficient for SWV was 0.563 and 0.605 for YM . However, MAS is often affected by factors, such as an examiner's experience, subjective individual differences, and understanding of the scale standards. It cannot accurately quantify changes in muscle tension, because an increase in muscle tension involves not only resistance felt by the limbs but also physical tissue properties and elasticity inside the muscle or connective tissue. Clinical evaluations of muscle tension are related only to the classification of resistance felt by the flexion and extension of the limbs. To date, there are no relevant data pertaining to specific changes in the elasticity inside the muscle or connective tissue and their relationship with classification of increased muscle tone [The diagnostic assessment of spasticity is based on the experience of a clinically evaluated physician or physical therapist. The MAS is the most commonly used assessment scale. It is the most widely used method in clinical practice, because this scale is easy to evaluate and does not require additional equipment and technical processing. The present study used MAS to evaluate the curative effect before and after rehabilitation of the upper limb spasm and to reflect the effect of rehabilitation treatment . There wcle tone .E = 3pc2 . The harder the structure is, the greater its density is, and the faster the SWV is, the higher its YM is [P < 0.01).Acoustic pulses emitted by the ultrasound probes are focused at different depths in the tissue to generate transverse shear waves. According to the Mach cone principle –15, a quts YM is . In the P > 0.05). After a stroke, a decrease in muscle fibers, cross-sectional muscle area, and proliferation of connective tissue around the muscle will lead to changes in muscle stiffness [The structural characteristics of muscles determine their biomechanical properties and are important determinants of muscle function . In the tiffness . By comptiffness  found thtiffness . AccordiP < 0.01). However, SWV and YM were still higher compared to the normal side after the rehabilitation treatment. SWV in the normal biceps brachii was 4.34 ± 1.32 m/s and 4.95 ± 1.02 m/s in the spastic biceps brachii after the rehabilitation treatment. YM in the normal biceps brachii was 50.58 ± 10.54 kPa and 72.08 ± 9.86 kPa in the spastic biceps brachii after the rehabilitation treatment. There was a statistically significant difference in SWV and YM in the spastic biceps brachii after the rehabilitation treatment (P < 0.05), which may be due to the short duration of the study. It is possible that SWV and YM in the biceps brachii on the spastic side may be similar to those on the normal side. Nevertheless, a decrease in SWV and YM indicates that the degree of muscle spasm was improved and that rehabilitation treatment was effective. In the present study, SWV and YM in the spastic biceps brachii were correlated with MAS before and after the rehabilitation treatment, with SWV and YM correlation coefficients of 0.563 and 0.605, respectively (P < 0.05). These data provide more objective evidence for rehabilitation physicians to determine whether the patient rehabilitation treatment is effective.In the present study, both SWV and YM in the biceps brachii were decreased after the rehabilitation treatment compared to those before rehabilitation. SWV in the spastic biceps brachii was 6.58 ± 1.16 m/s, and YM was 123.34 ± 25.74 kPa before the rehabilitation treatment. After rehabilitation, SWV in the spastic biceps brachii was 4.95 ± 1.02 m/s and YM was 72.08 ± 9.86 kPa. There was a statistically significant difference in SWV and YM in the spastic biceps brachii before and after the rehabilitation treatment (In conclusion, SWE can be used as a quantitative index to analyze changes in the muscle structure and function after stroke and as a means of assessment before and after the rehabilitation treatment.


Pulmonary rehabilitation offers potential benefits to people with asthma. It is however unknown if rehabilitation favourably affects patients’ health care needs. We therefore examined if rehabilitation reduced needs and, in addition, if it improved asthma control.One hundred fifty patients with asthma were surveyed in three rehabilitation clinics at admission and at discharge. Additionally, we surveyed 78 participants with asthma twice 4 weeks apart. The latter sample (i.e. the control group) was recruited through other pathways than rehabilitation clinics. The Patient Needs in Asthma Treatment (NEAT) questionnaire and the Asthma Control Test (ACT) were completed at baseline and follow-up. Differences between baseline and follow-up and between rehabilitation and control group were examined by t-tests and chi-squared-tests. Univariate ANCOVAS were used to examine if NEAT and ACT follow-up scores differed significantly between groups. Within the rehabilitation group, linear regressions were used to examine if self-reported utilization of more interventions that addressed needs were associated with NEAT scores at follow-up.t(149) = 10.33, p <  0.01) and increased asthma control (t(130) = -6.67, p <  0.01), whereas members of the control group exhibited no changes. Univariate ANCOVAS showed that unmet follow-up needs  = 36.46, p <  0.001) and follow-up asthma control  = 6.97, p = 0.009) differed significantly between groups. In the rehabilitation group, self-reported utilization of more interventions was associated with reduced needs .At baseline, there were no differences between the rehabilitation and the control group regarding needs and asthma control. At follow-up, the rehabilitation group showed reduced needs (This study provides preliminary evidence suggestion that pulmonary rehabilitation in adults with asthma may reduce asthma-related needs and confirms previous findings that rehabilitation may improve asthma control. Pulmonary rehabilitation is considered a key factor in the successful long-term care of patients suffering from asthma. This holds particularly true whenever patients experience adverse physical, social or psychological consequences due to their asthma despite access to and availability of adequate outpatient medical care . PulmonaGuidelines on pulmonary rehabilitation –6 and reneed should not be understood as an objective need or a need recommended by guidelines or the physician (i.e. normative need), but instead as a perceived health care need that is felt by a given patient and remains unmet (e.g. information or support needs) [Patient-centeredness in asthma care also includes consideration of patients’ health care needs. Addressing unmet health care needs may improve patients’ adherence and health outcomes such as asthma control , 20. In t needs) , 22. To t needs) . The NEAt needs) , 18.In order to examine if pulmonary rehabilitation can improve patients’ health care needs and asthma control, we collected data among two samples. First, we included patients who participated in an inpatient rehabilitation with specific asthma-related interventions (rehabilitation group). Second, to facilitate a comparison, we included a control group.Between August 2018 and January 2019, we carried out a survey among 150 people undergoing pulmonary rehabilitation in three rehabilitation clinics in Germany, all of which adopted a bio-psycho-social approach with individualized interventions (i.e. selection of treatment according to patients’ needs). All people were at least 18 years old, German speaking, and had physician-diagnosed asthma, but did not have COPD. Participants were recruited through attending physicians during admission and, if inclusion criteria were met, were asked to complete the survey (by paper-and-pencil) at admission (before the treatment period) and at discharge . This means that the time between baseline and follow-up differed between participants, depending on the duration of rehabilitation. All patients filled in the questionnaire at the respective rehabilitation clinic. Participants in the rehabilitation group received the usual scope of care that is common in the respective rehabilitation clinic.Between January and July 2018, we carried out a survey among 78 adults (≥18 years) who reported physician-diagnosed asthma. Participants were recruited by various pathways, e.g. by support groups or pulmonary sport groups, and were asked to complete the questionnaire twice (either web-based or by paper-and-pencil) 4 weeks apart. The participants did not have to meet any asthma-related inclusion criteria (e.g. being stable) and could fill in the questionnaire at home. The data from this sample has already been published elsewhere [Unmet needs were captured by the NEAT questionnaire , 24. ThiIn addition to health care needs, we assessed asthma control, measured by the well-established Asthma Control Test (ACT). The ACT is a 5-item instrument assessing asthma symptoms, use of rescue medications, and the effect of asthma on daily functioning . The ACT’s potential score ranges from 5 (very poorly controlled) to 25 (completely controlled) with higher scores indicating better control. A score of 19 or less has been defined as a cut-off score suggesting poor control .We examined to what extent participants in the rehabilitation group received specific asthma treatment or support interventions during rehabilitation that may be assumed to exert effects on specific domains on health care needs. Therefore, we asked for asthma education and training that reflected needs, surveyed in the NEAT questionnaire. In detail, we asked rehabilitation group participants the following question: “What content was conveyed during asthma education?” Response options were based on a tick-off list, e.g. “information about how to take my asthma drugs” or “practical exercise of breathing techniques, which can help during asthma attack”. We collected this data to be able to examine if (specific) PR interventions were associated with reduced needs in a dose-response fashion.In order to describe and compare both samples appropriately, we present baseline data on gender, age, school education, reported allergy (based on a tick-off list) and follow-up time.Whereas health care needs, asthma control and demographic information were gathered in the rehabilitation and the control group, questions on specific asthma education and training during rehabilitation were only asked in the rehabilitation group.Initially, comparisons between rehabilitation and control group regarding demographics as well as NEAT and ACT scores were carried out by unpaired t-tests  and chi-squared-tests  to examine if the samples’ measured characteristics were sufficiently comparable.In both samples, we examined the potential changes of the NEAT total scores and subscale-specific scores by pre-post-score comparisons and tested for statistical significance based on paired t-tests. In addition, we examined the respective potential changes of the ACT sum scores also by running paired t-tests. We hypothesized that the NEAT scores would decline (i.e. indicating reduced needs) and that the ACT scores would increase (i.e. reflecting better asthma control) during the follow-up period only in the rehabilitation, but not in the control group.Furthermore, we ran univariate ANCOVAS with group membership  as independent variable and NEAT and ACT follow-up scores as dependent variables. To consider the potentially dissimilar demographic characteristics of the rehabilitation and the control group, we included gender, age, educational level, number of allergies, follow-up period and the respective baseline score (NEAT or ACT) as covariates.exacerbations was expected; however, if a patient reported not to have learned about patient-physician communication, a reduction of the NEAT patient expertise subscale score was not to be expected. Linear regressions were controlled for gender, age, educational level, duration of stay as well as for a variable reflecting the three different rehabilitation clinics.Finally, to examine if pulmonary interventions were associated with reduced health care needs in a dose-response fashion, we ran linear regression models with the total sum score of self-reported received asthma education/training during rehabilitation that addressed needs as independent variable and the NEAT total change score as dependent variable. Furthermore, we calculated subscale-specific scores capturing the type of received asthma education/training, which reflect subscale-specific topics of the NEAT. For instance, if breathing techniques and correct use of the inhaler were taught to a given patient, a reduction of the corresponding NEAT score on the subscale In the rehabilitation group, 178 participants completed the baseline questionnaire at admission and 166 filled in the follow-up questionnaire at discharge (response rate = 93%). We excluded an additional 16 participants either because they completed the baseline questionnaire more than 3 days after admission and had therefore possibly received some interventions prior to that or because they filled out the follow-up questionnaire more than 3 days before discharge and interventions were thus potentially still pending. The final sample for our analysis comprised thus 150 participants.In the control group, 112 people with asthma participated in the first assessment and 78 of those provided data on both occasions (response rate = 67%).Table p <  0.001), age , school education , reported allergies  and follow-up period .Samples differed significantly in terms of gender . This was explained by a significant reduction of unmet needs in the rehabilitation group  = 10.33; p <  0.001) compared to their baseline score whereas the score in the control group remained stable . In contrast, at follow-up, we found a significantly lower NEAT total mean score in the rehabilitation as compared to the control group . We found that in the rehabilitation group asthma control significantly increased during follow-up period to an rather appropriate level (well controlled ≥ 20) (t(130) = − 6.67, p <  0.001), whereas in the control group asthma control remained poor and exhibited little and non-significant change . In contrast, at follow-up, the rehabilitation group showed significantly higher ACT mean scores compared to the control group  = 36.46, p <  0.001), and the level of asthma control at follow-up  = 6.97, p = 0.009), differed significantly between the two groups . These observations imply that the rehabilitation group showed significantly less health care needs and better asthma control at follow-up (and after pulmonary rehabilitation) compared to the control group.Univariate ANCOVAS Table  showed tp = 0.03) as well as measured by the change scores of the NEAT subscales patient expertise  and exacerbations . Further, received interventions that addressed patient expertise were significantly associated with reduced needs, measured by the change score of the NEAT subscale patient expertise  and received interventions addressing exacerbations tended to be significantly associated with reduced needs, measured by the change score of the NEAT subscale exacerbations . In contrast, there was no evidence of associations between received interventions addressing drug effects and the change score of the NEAT subscale drug effects as well as between experienced interventions addressing how to handle drugs and the change score of the NEAT subscale handling drugs.In the rehabilitation group we ran additional linear regression models to examine if self-reported utilization of more interventions that address needs, surveyed in the NEAT questionnaire, were associated with reduced health care needs in a dose-response fashion Table . We founpatient expertise . Also, received interventions addressing exacerbations were significantly associated with reduced needs, measured by the change score of the NEAT subscale drug effects  and received interventions addressing treatment in drug effects were significantly associated with reduced needs, measured by the change score of the NEAT subscale exacerbations .Furthermore, received interventions addressing education in handling drugs were significantly associated with reduced needs, measured by the change score of the NEAT subscale This study provides initial evidence suggestion the benefits of pulmonary rehabilitation in adults with asthma in relation to health care needs and confirms prior research on asthma control.patient expertise. These findings suggest that not only a better exchange of information between patient and physician, but also a more appropriate handling of drugs may reduce the need for being an expert for one’s own disease. Furthermore, received interventions addressing topics like information on what to do in case of an asthma attack or practical training of breathing techniques tended to be significantly associated with less health care needs in the NEAT subscale exacerbations. Similarly, information on drug effects, side effects and interactions of drugs led to reduced health care needs regarding exacerbations. Thereby, we can assume that not only patient education referring to what to do in case of an exacerbation, but also better knowledge of drug effects may help to reduce needs regarding information on what to do in case of exacerbations. Also, received interventions addressing exacerbations were associated with reduced health care needs in the NEAT subscale drug effects. Thereby we can assume that more skills and knowledge regarding possible asthma attacks may lead to a decrease in health care needs regarding drug effects.Initially, we found that unmet health care needs were reduced during rehabilitation and that this effect was associated with the utilization of suitable asthma interventions in a dose-response fashion. To our knowledge, there exist no studies examining the association between pulmonary rehabilitation and (unmet) health care needs and we therefore provide novel evidence. To gain a better understanding, we additionally calculated subscale specific scores on received asthma education/training, which reflect subscale specific topics of the NEAT. Our results indicate that better physician-patient-communication, e.g. physician made more time available in case of special requests or considered personal circumstances during treatment, but also practical training and information on how to use asthma drugs were associated with less health care needs regarding the NEAT subscale Given the findings from our previous study on psychometric properties , we haveIn summary, we can assume that a) interventions during rehabilitation are associated with reduced health care needs in a dose-response fashion and b) very specific treatment elements may reduce specific health care needs during rehabilitation. Compared to the rehabilitation group, we did not find any improvement of health care needs in patients who most likely did not receive any kind of new intervention (control group). By using ANCOVA we were able to ensure that this result is due to participation in rehabilitation and not merely to demographic differences such as age, gender or educational level. Results of the ANCOVA showed that members of the control group had significant more health care needs at follow-up than members of the rehabilitation group. Thus, one can be assumed that unmet health care needs in the rehabilitation group did not decrease merely due to demographic characteristics.Based on these results, our study should be understood as an initial step to provide evidence. Experimental studies that are based on the randomized allocation of pulmonary rehabilitation and that maximize the likelihood of adequate control groups  are needed to provide more valid insights into the causal relations between rehabilitation and health care needs. Although conducting an RCT on this matter remains challenging, as people with considerable health problems cannot be withheld from rehabilitation.F = 44.65, p <  0.001). This finding could indicate that especially the latter could benefit from rehabilitation.To our knowledge, there is only one waiting-list RCT of pulmonary rehabilitation in patients with uncontrolled asthma and this RCT demonstrated that rehabilitation is effective, e.g. in terms of asthma control . This stThe improvement of asthma control between baseline and follow-up was not observed in adults with asthma who most likely did not receive any kind of new intervention during follow-up period: control remained poor in this sample. Building on this finding, we calculated a univariate ANCOVA , which also showed that members of the control group had significantly less asthma control at follow-up than members of the rehabilitation group. Overall, our results regarding asthma control support previous findings on the impact of pulmonary rehabilitation on patients’ health care , 18.residual confounding) cannot be excluded. Furthermore, unfortunately no information on medication was collected. Therefore, we are unable to examine a possible link between pulmonary rehabilitation and medication (e.g. reduction of doses).Importantly, our results are not based on data from an RCT, but we employed a quasi-experimental design. A RCT is considered the gold standard to examine effectiveness of interventions and its defining feature is randomization. Randomization implies that participants are randomly allocated to either the intervention or the control group which should, if the trial is sufficiently large, yield two samples that are comparable in terms of measured and unmeasured confounders . A quasiIt is reassuring though that there were no baseline differences in terms of our primary outcomes (NEAT and ACT) between rehabilitation and control group. Furthermore, despite using age, gender educational level, number of allergies and follow-up period as covariates, members of the control group had significant more health care needs and less asthma control at follow-up than members of the rehabilitation group. Also, we included individuals from three rehabilitation clinics and thereby collected data in a multicentre design. Thus, we can assume that the improvement of health care needs and asthma control is not limited to an intervention in a specific clinic (i.e. single centre study); rather it seems that rehabilitation by itself and regardless of the clinic or its slightly different style of intervention administration may improve these outcomes.Unfortunately, we cannot know with certainty that participants in the control group did not receive any new intervention (like elements of a disease management program) during the 31-day-follow-up period. However, since the period was short, we can assume that no rehabilitation took place. Furthermore, unfortunately no information on medication was collected. Therefore , we are unable to examine a possible link between pulmonary rehabilitation and medication (e.g. reduction of dose).p = 0.03). Also, there existed no significant differences between responders and non-responders in both samples regarding NEAT and ACT scores at baseline. In summary, we can assume that major selection bias across the follow-up is rather unlikely.A special strength of both samples is that we attained good response rate at follow-up . Furthermore, responders and non-responders provided similar background data at baseline, except for reported allergies in the control group, which were more common in responders  to evaluate the interventions and to examine if important cornerstones are met from patients’ point of view.The NEAT has been found to be a reliable and valid instrument to predict treatment satisfaction  and therThis study provides preliminary evidence suggestion of the benefits of pulmonary rehabilitation in adults with asthma in reducing patients’ unmet health care needs and confirms previous findings that rehabilitation may improve asthma control.


To identify the genetic cause of complex neuropathy in two siblings from a consanguineous family.The patients were recruited from our clinic. Muscle biopsy and whole-exome sequencing (WES) were performed. Fibroblasts cell lines from the index patient, unaffected parents, and three normal controls were used for cDNA analysis and western blot.SBF1 gene in the two siblings, while both parents and the unaffected sibling were heterozygous carriers. Functional analysis showed a markedly reduced level of MTMR5 protein encoded by SBF1 in the index case. The levels of MTMR5 protein in unaffected parents were similar to those found in controls.The index patient was a 29-year-old male with clinical phenotype of syndactyly, pes cavus, swallowing difficulties, vision problem, imbalance, and muscle weakness. The sibling had similar, but milder symptoms. Nerve conduction studies and electromyography of both patients suggested sensory-motor axonal neuropathy. Muscle biopsy showed a feature of necklace fibres. WES identified a novel homozygous frameshift deletion  in exon 40 of the SBF1 was identified in this family. Sensory-motor axonal neuropathy and necklace fibres in biopsy were the major features expanding the phenotypic spectrum of SBF1-related recessive syndromic neuropathy.A novel homozygous frameshift deletion in The online version of this article (10.1007/s00415-020-09827-y) contains supplementary material, which is available to authorized users. MTMR2 (OMIM#603557) [MTMR13/SBF2 (OMIM#607697) [MTMR5/SBF1 (OMIM#603560) [SBF1 have also been reported in four families with severe syndromic manifestations such as sensory-motor polyneuropathy, multiple cranial nerve involvement, cognitive delay, and skeletal abnormalities [SBF1 was identified in two siblings with complex neuropathy and “necklace fibres” in muscle biopsy, expanding the spectrum of SBF1-related syndromic neuropathy.Charcot–Marie–Tooth (CMT) disease is a large group of clinically and genetically heterogeneous inherited neuropathies which are characterised by slowly progressive distal muscle weakness, wasting, and sensory loss. Autosomal recessive forms of demyelinating CMT are also called CMT4 and a specific subgroup CMT4B is characterised by severe demyelinating neuropathy with focal myelin outfoldings and secondary axonal loss . Mutatio2 OMIM#60557 [MTMRThe affected siblings from a British family  at the age of two, and had a history of photophobia dating back to the age of three. Over the years, the patient developed symptoms of slowness of swallowing, choking on food, blurred and double vision, imbalance, dragging his legs, easy fatigue, and weakness. These symptoms, particularly his gait, speech, and eyelid drop, were worse in the evening. The patient currently wears ankle foot to support his walking. On examination, he had obvious strabismus, gaze-evoked nystagmus to the left, as well as on up-gaze, and he had broken pursuit movements. He has a tonic pupil not reactive to light or accommodation. His fundi were normal. The rest of cranial nerve examination was normal. Power in lower limbs was Grade 4/5 throughout. Distal weakness in wrist extension was at 4/5, and finger extension and flexion were at 3/5. He displayed symmetrical hypo-reflexia with bilateral extensor plantars; however, his coordination was normal. Sensory examination showed that he had a patchy pin-prick loss in a glove and stocking distribution. Blood tests were negative for acetylcholine receptor antibodies, anti-MUSK antibody, and anti-voltage gated calcium channel antibody. The creatine kinase level and metabolic testing were normal. Electromyography and nerve conduction study of both patients were performed, and showed moderately severe axonal sensory-motor neuropathy (Table The index patient II-2) and his affected sister II-1) were from a consanguineous family where parents are second cousins  of the index patient also had similar pupils and disc appearance to the index patient, with slightly difficult walking, some weakness and wasting in her hands and feet, and endometriosis. On examination, her eyes showed slightly jerky pursuit movement. She had thin wrists and small hands but no clear wasting and only slight weakness of the thumbs. Her feet were similar to the hands, with only minimal weak dorsiflexion. She had similar sensory loss in a glove and stock distribution, and her feet also became discoloured, often black and quite cold. Fig. h–j She aBoth of their parents and the younger brother were not affected. Overall, the phenotype in this family is consistent with an autosomal recessive form of inheritance.SBF1/MTMR5 gene, a known gene for CMT. This deletion is absent in the above public databases. The variant were confirmed by Sanger sequencing. Segregation analysis in both parents and two siblings found that the affected sister (II-1) also harbours the same homozygous frameshift deletion, and both parents as well as the unaffected sibling are heterozygous carriers of the deletion in SBF1  < 1% in the 1000 Genomes project, Exome Variant Server, and the Exome Aggregation Consortium (ExAC) databases. In the filtering steps, we also prioritised for homozygous or compound heterozygous variants in autosomal recessive genes. This left a novel homozygous frameshift deletion  in exon 40 of the SBF1 (Supplementary Table). They share several similar phenotypes with previously reported families with CMT4B3. Ophthalmoplegia was seen in most reported cases in addition to progressive weakness of the limbs and sensory-motor axonal neuropathy [SBF1-related cases. Necklace fibres with nuclear internalisation have been reported as a histological hallmark of X-linked myotubular myopathy related to mutations in the phosphoinositide phosphatase myotubularin 1 (MTM1) gene and also a late-onset dynamin-2 (DNM2) gene-related centronuclear myopathy [MTM1 nor DNM2 was found in the index patient. Our study is the first to report necklace fibres in SBF1-related phenotypes, which expands the clinical and molecular genetic spectrum of this specific pathological feature.Our family is the sixth family identified with mutations in uropathy , 9. Deveuropathy , 7, 10, myopathy , 12. Themyopathy . NeitherSBF1 gene encodes protein MTMR5, a member of the myotubularin-related (MTMR) family. MTMR proteins are a large family of phosphoinositide (PI) 3-phosphatases which are involved in membrane trafficking and endocytosis [The ocytosis . The proocytosis . Some meocytosis . Some meocytosis . Studiesocytosis .SBF1 are linked to male infertility [SBF2 gene, leads to alterations in MTMR2 function and subsequent dysfunction of membrane trafficking, endocytic/exocytic processes, and neuron–Schwann cell interactions [Loss of function of MTMR5 was suggested to cause a defect in the late stages of spermatogenesis in mice  and studertility . Howeverractions . MTMR13 ractions . The levractions . MTMR2 aractions . This coMTMR2, SBF2, and SBF1 genes.As MTMR5 is the closest homologue of MTMR13, it is possible that the loss of function in MTMR5 could also induce dysfunction in neuronal or Schwann cells via a similar mechanism with the MTMR2/MTMR13 complex. This may explain the strikingly similar phenotypes of patients with recessive mutations in the SBF1 identified in both affected siblings, indicating a loss-of-function mechanism underlying the pathogenesis of the disease. Whether this causes the patient to be infertile is as yet unknown.In the present study, the protein level of MTMR5 was markedly reduced due to the homozygous frameshift deletion in SBF1 in a family affected with a rare form of neuropathy with necklace fibres on the muscle biopsy. Segregation and functional analysis suggested that the homozygous deletion in SBF1 was the main culprit to the disease via a loss-of-function mechanism. Our findings in this family expand the phenotypic spectrum of SBF1-related recessive syndromic neuropathy.We reported a novel homozygous frameshift deletion in Supplementary file1 (DOCX 25 kb)Below is the link to the electronic supplementary material.


The Excellence in Rehabilitation of Aging Persons Award lecture will feature an address by the 2020 Excellence in Rehabilitation Award recipient Kenneth J. Ottenbacher, PhD, OTR. The Excellence in Rehabilitation of Aging Persons Award is designed to acknowledge outstanding contributions in the field of the rehabilitation of aging individuals.