Rabdomiossarcoma cutâneo primário de padrão alveolar: relato de caso em adulto jovem / Primary cutaneous alveolar rhabdomyosarcoma: Case report in a young adult

Sarcomas são neoplasias mesenquimais malignas, raras, que acometem, principalmente, crianças e adolescentes. O rabdomiossarcoma, subtipo oriundo da musculatura esquelética, é condição incomum em adultos, acometendo sítios de localização não habitual, crescimento rápido e de difícil tratamento. Apresenta-se caso de adulto jovem com nodulação em lóbulo auricular esquerdo, cuja análise histopatológica e imuno-histoquímica confirmou tratar-se de rabdomiossarcoma alveolar, o qual foi conduzido em conjunto com a Oncologia.

Sarcomas are rare and malignant mesenchymal neoplasms that mainly affect children and adolescents. Rhabdomyosarcoma, a subtype originating from skeletal muscle, is an uncommon condition in adults. It affects sites of unusual location, presents fast growth, and is challenging to treat. We report a case of a young adult with nodules in the left auricular lobe. The histopathological and immunohistochemical analysis confirmed the alveolar rhabdomyosarcoma, and treatment was conducted in association with Oncology.

Lack of dystrophin influences muscle inflammation but not myogenic regulatory factors after eccentric exercise in mdx mice

Abstract Aim: To investigate the consequences of chronic eccentric exercise in histopathology, inflammatory, and myogenic regulatory factors response in gastrocnemius muscle of X-chromosome-linked muscular dystrophy (mdx) mice. Method: Male mdx and control mice (C57BL/10 lineage) were distributed in the following groups: Sedentary Control (SC), Trained Control (TC), Sedentary Mdx (S-Mdx), and Trained Mdx (T-Mdx). Trained animals were subjected to downhill running for 7 weeks. Gastrocnemius was submitted to histopathological analysis and immunoexpression of Cyclooxygenase-2 (COX-2) and myogenic regulatory factors (myoD and myogenin). Results: The exercise influenced inflammation response as demonstrated by the increased COX-2 immunoexpression in T-Mdx. Interestingly, Myogenic regulatory factors revealed that the lack of dystrophin has not been influenced myoD and the increase of myogenin occurred due to exercise and was not aggravated by the absence of dystrophin. Conclusion: In conclusion, an eccentric exercise in gastrocnemius of mdx mice was characterized by an intense inflammatory process without myogenic response. These findings suggest that special attention should be given to inflammatory aspects related to COX-2 associated with a decrease of myoD expression, as biomarkers in motor rehabilitation programs.

Effect of electroacupuncture on tissue elasticity and expression of muscular MyoD and myogenin in rats with acute gastrocnemius contusion / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Ashi" acupoint and "Kunlun" (BL60) on elastic modulus, histopathological changes and expression of myogenic regulatory factors in gastrocnemius(GM) contusion rats, so as to explore the therapeutic effect of local acupoint selection and acupoint selection along channel. METHODS: Male SD rats were randomly divided into blank control (n＝5), model (n＝15), Ashi-point (n＝15) and BL60 (n＝15) groups. The acute GM contusion model was established by striking (free falling) the GM with a homemade hitter. EA (0.5 to 1.0 mA, 2 Hz/10 Hz) was applied to Ashi-point (local focus) and BL60 for 30 min 24 h after muscle injury. The elasticity maximum (Emax) of gastrocnemius muscle was measured by using an ultrasonic device. Histopathological changes were observed after H．E. stain, and the number of Myogenic differentiation(MyoD)- and Myogenin (MyoG)-positive cells was detected by using immunohistochemistry. RESULTS: After mdeling, the Emax value of GM was significantly increased from the 3rd h to 7th day in comparison with pre- injury of muscle (P<0.05), and was markedly increased on the 3rd day and obviously lower on day 7 in the Ashi-point group than in the model group (P<0.05). The numbers of MyoD- and MyoG-positive cells of GM were significantly increased on day 7 in the model group than in the blank control group (P<0.05), and both further increased in Ashi-point on day 3 and 5, and MyoG-positive cells further increased in BL60 group on day 5 and in Ashi-point group on day 7 relevant to the model group(P<0.05). The therapeutic effect of EA-Ashi-point was apparently superior to that of BL60 in up-regulating Emax on day 3 and in up-regulating the number of MyoD-positive cells on day 3 and 5 (P<0.05). H．E. stain showed disordered arrangement of muscle fibers, infiltration of inflammatory cells, increase of intercellular space, and edema on day 3 after modeling (which was milder in the Ashi-point group); and gradual fusion and thickening of new born muscle fibers with obvious connective tissue hyperplasia converged to the lesioned region on day 7 in the model group (convergence of new born muscle cells to the lesion region in both EA groups, and more complete tissues in the Ashi-point group). CONCLUSION: EA of Ashi-point and BL60 can up-regulate the expression of myogenic regulatory factors MyoD and MyoG of GM tissue in GM contusion rats, which may contribute to its function in promoting recovery of muscle elasticity. The role of EA-Ashi-point is superior to that of EA-BL60.

Effect of electroacupuncture of acupoints on the healthy limb (opposing needling) on acute skeletal muscle contusion in rats / 针刺研究

OBJECTIVE: To observe the therapeutic effect of electroacupuncture (EA) of "Zusanli" (ST36) and "Ashi"-point on the healthy side (opposing needling) on muscular injury and expression of myogenin (myoG) and fast myosin skeletal heavy chain (Fast MyHC) proteins in the gastrocnemius muscle (GM) tissues in skeletal muscle contusion rats，so as to explore its mechanism underlying improvement of skeletal muscle injury. METHODS: A total of 54 male SD rats were divided into normal control (n ＝ 6)，model (n＝24) and opposing needling (EA, n＝24) groups. The latter two groups were further randomized into 3, 5, 7 and 14 d subgroups (n＝6 per subgroup). The skeletal muscle contusion model of the hind-limb was established by using a self-made striking device. EA (1 Hz/3 Hz，1－2 mA) was applied to ST36 and "Ashi"-point on the uninjured side of the hind-limb for 15 min every time, once a day for 3, 5, 7 and 14 days, respectively. The injured GM was harvested on the 3rd, 5th, 7th and 14th day after muscular contusion. The morphological changes of the injured GM and the mean cross-sectional areas (CSAs) of the neonatal muscle cells were observed by microscope after H．E. staining. The immunoactivity of desmin protein (myogenic marker protein of myoblast cell) of GM was detected by immunofluorescence stain on the 7th day after injury, and the expression levels of myoG (on the 3rd and 5th day after injury) and fast MyHC protein of GM tissues (on the 7thand 14th day after injury) were detected by Western blot. RESULTS: H．E. staining of GS tissue showed fewer neuronal myocytes with disordered arrangement at different sizes, and appearance of some collagenous fibers among the mesenchyme on day 7 and 14 after muscular contusion, which was relatively milder in the EA group. In the EA group, the CSA values of the neonatal muscle cells were significantly larger than those in the model group on the day 7th (P0.05). On the 3rd and 5th day after muscular contusion, the expression level of myoG protein was significantly up-regulated in the model group compared with the normal control group (P<0.001), and significantly up-regulated in the EA group than that in the model group (P<0.001). On the 7th and14th day after contusion, the expression level of fast MyHC protein was significantly down-regulated in the model group relevant to the normal control group (P<0.001), and markedly up-regulated in the EA group relevant to the model group (P<0.01)．. CONCLUSION: EA of ST36 and "Ashi"-point on the contralateral limb can up-regulate the expression of myoG and fast MyHC proteins of GM in acute skeletal muscle contusion rats, which may contribute to its effect in promoting the repair of skeletal muscle injury.

Rheumatoid arthritis-increased gene expressions in muscle atrophy are restored back to control as a response to acute resistance exercise / Estímulo agudo de musculação promove regulação de genes alterados pela atrofi a muscular

This study had as objective to analyze the acute eff ects of resistance exercise (RE) on the mRNA levels of the following genes (MyoD, myogenin, IGF-1, atrogin-1, MuRF-1, and myostatin) in rheumatoid arthritis (experimental arthritis). Therefore, 26 females rats were randomly allocated into four groups, control (CT, n=7), exercise (Ex, n=6), rheumatoid arthritis (RA, n=6) and RA with exercise (RAEx, n=7). Met-BSA was injected into the tibiotarsal joint in the RA and RAEx groups. After 15 days from injection, the animals were submitted to an acute bout of RE and six hours post protocol the animals were euthanized. We evaluated the joint thickness, infl ammation score, cross-sectional area (CSA) of gastrocnemius muscle fi bers and mRNA expression of the IGF-1, MyoD, myogenin, myostatin, MuRF-1, atrogin-1 and GAPDH. It was observed that the joint thickness and score strongly increased in arthritic rats (p <0.001) while the CSA decreased (p ≤ 0.05). Increased mRNA levels of IGF-1 (2.0 fold), myostatin (4.5 fold), atrogin-1 (2.5 fold), MyoD (3.7-fold) and myogenin (5 fold) were observed in muscle of arthritic rats. The mRNA expression of myostatin, atrogin-1, MyoD and myogenin decreased in the RAEx group. In this way, we can conclude that experimental arthritis-increased gene expressions in muscle atrophy myostatin, atrogin-1, MyoD and myogenin) are restored back to control as a response to acute RE....(AU)

O presente estudo teve como objetivo analisar o efeito agudo do Exercício com pesos sobre os níves de mRNA de genes envolvidos no anabolismo ou catabolismo muscular em um modelo experimental de Artrite Reumatóide. Para tanto, 26 ratas fêmeas foram randomicamente alocadas em quatro grupos, controle (CT, n=7), Exercício (Ex, n=6), Artrite Reumatóide (AR, n=6) e Artrite Reumatóide com exercício (AREx, n=7). Uma substância contendo Albumina bovina metilada foi injetada na articulação tíbio-tarsal nos grupos AR e AREx para indução da Artrite Reumatóide. Após 15 dias da injeção, os animais foram submetidos a um estímulo agudo de treinamento com pesos e 6 horas após o exercício os animais foram eutanasiados. Nós avaliamos a espessura da articulação, escore de infl amação, a área de secção transversa (AST) das fi bras do músculo Gastrocnêmio e a mRNA de IGF-1, MyoD, Myogenina (genes envolvidos no anabolismo muscular), e MuRF-1, atrogina-1 (genes envolvidos no catabolismo muscular), além do gene controle , GAPDH. Foi observado que a espessura articular e o escore de infl amação aumentaram fortemente nas ratas induzidas a Artrite Reumatóide (p <0,001), enquanto a AST reduziu (p ≤ 0,05). Um aumento nos níveis de mRNA de IGF-1 (2,0 vezes), miostatina (4,5 vezes), atrogina-1 (2,5 vezes), MyoD (3,7 vezes) e miogenina (5 vezes) foi observado no músculo das ratas induzidas a Artrite Reumatóide. mRNA de miostatina, atrogina-1, MyoD e miogenina reduziu no grupo RAEx. Desta forma, podemos concluir, que o modelo experimental de Artrite Reumatóide induziu um aumento da expressão de genes durante a atrofi a muscular (myostatin, atrogin-1, MyoD and myogenin) e que estas alterações foram reguladas pelo Exercício com peso....(AU)

Implantation of muscle satellite cells overexpressing myogenin improves denervated muscle atrophy in rats

This study evaluated the effect of muscle satellite cells (MSCs) overexpressing myogenin (MyoG) on denervated muscle atrophy. Rat MSCs were isolated and transfected with the MyoG-EGFP plasmid vector GV143. MyoG-transfected MSCs (MTMs) were transplanted into rat gastrocnemius muscles at 1 week after surgical denervation. Controls included injections of untransfected MSCs or the vehicle only. Muscles were harvested and analyzed at 2, 4, and 24 weeks post-transplantation. Immunofluorescence confirmed MyoG overexpression in MTMs. The muscle wet weight ratio was significantly reduced at 2 weeks after MTM injection (67.17±6.79) compared with muscles injected with MSCs (58.83±5.31) or the vehicle (53.00±7.67; t=2.37, P=0.04 and t=3.39, P=0.007, respectively). The muscle fiber cross-sectional area was also larger at 2 weeks after MTM injection (2.63×103±0.39×103) compared with MSC injection (1.99×103±0.58×103) or the vehicle only (1.57×103±0.47×103; t=2.24, P=0.049 and t=4.22, P=0.002, respectively). At 4 and 24 weeks post-injection, the muscle mass and fiber cross-sectional area were similar across all three experimental groups. Immunohistochemistry showed that the MTM group had larger MyoG-positive fibers. The MTM group (3.18±1.13) also had higher expression of MyoG mRNA than other groups (1.41±0.65 and 1.03±0.19) at 2 weeks after injection (t=2.72, P=0.04). Transplanted MTMs delayed short-term atrophy of denervated muscles. This approach can be optimized as a novel stand-alone therapy or as a bridge to surgical re-innervation of damaged muscles.

Effect of low-level laser therapy (808 nm) on skeletal muscle after endurance exercise training in rats

BACKGROUND: Low-level laser therapy (LLLT) has been demonstrated to be effective in optimizing skeletal muscle performance in animal experiments and in clinical trials. However, little is known about the effects of LLLT on muscle recovery after endurance training. OBJECTIVE: This study evaluates the effects of low-level laser therapy (LLLT) applied after an endurance training protocol on biochemical markers and morphology of skeletal muscle in rats. METHOD: Wistar rats were divided into control group (CG), trained group (TG), and trained and laser irradiated group (TLG). The endurance training was performed on a treadmill, 1 h/day, 5 days/wk, for 8 wk at 60% of the maximal speed reached during the maximal effort test (Tmax) and laser irradiation was applied after training. RESULTS: Both trained groups showed significant increase in speed compared to the CG. The TLG demonstrated a significantly reduced lactate level, increased tibialis anterior (TA) fiber cross-section area, and decreased TA fiber density. Myogenin expression was higher in soleus and TA muscles in both trained groups. In addition, LLLT produced myogenin downregulation in the TA muscle of trained animals. CONCLUSION: These results suggest that LLLT could be an effective therapeutic approach for stimulating recovery during an endurance exercise protocol.

Potential gene regulatory role for cyclin D3 in muscle cells.

Cyclin D3 is important for muscle development and regeneration, and is involved in post-mitotic arrest of muscle cells. Cyclin D3 also has cell-cycle-independent functions such as regulation of specific genes in other tissues. Ectopic expression of cyclin D3 in myoblasts, where it is normally undetectable, promotes muscle gene expression and faster differentiation kinetics upon serum depletion. In the present study, we investigated the mechanistic role of cyclin D3 in muscle gene regulation. We initially showed by mutational analysis that a stable and functional cyclin D3 was required for promoting muscle differentiation. Using chromatin immunoprecipitation assays, we demonstrated that expression of cyclin D3 in undifferentiated myoblasts altered histone epigenetic marks at promoters of muscle-specific genes like MyoD, Pax7, myogenin and muscle creatine kinase but not non-muscle genes. Cyclin D3 expression also reduced the mRNA levels of certain epigenetic modifier genes. Our data suggest that epigenetic modulation of muscle-specific genes in cyclin-D3-expressing myoblasts may be responsible for faster differentiation kinetics upon serum depletion. Our results have implications for a regulatory role for cyclin D3 in muscle-specific gene activation.

Effects of Sunphenon and Polyphenon 60 on proteolytic pathways, inflammatory cytokines and myogenic markers in H2O2-treated C2C12 cells.

The effect of Sunphenon and Polyphenon 60 in oxidative stress response, myogenic regulatory factors, inflammatory cytokines, apoptotic and proteolytic pathways on H2O2-induced myotube atrophy was addressed. Cellular responses of H2O2-induced C2C12cells were examined, including mRNA expression of myogenic regulatory factors, such as MyoD and myogenin, inflammatory pathways, such as TNF-α and NF-kB, as well as proteolytic enzymes, such as μ- calpain and m-calpain. The pre-treatment of Sunphenon (50 μg/mL)/Polyphenon 60 (50 μg/mL) on H2O2-treated C2C12 cells significantly down-regulated the mRNA expression of myogenin and MyoD when compared to those treated with H2O2-induced alone. Additionally, the mRNA expression of μ-calpain and m-calpain were significantly (p<0.05) increased in H2O2-treated C2C12 cells, whereas pre-treatment with Sunphenon/Polyphenon significantly down-regulated the above genes, namely μ-calpain and m-calpain. Furthermore, the mRNA expression of TNF-α and NF-kB were significantly increased in H2O2-treated C2C12 cells, while pre-treatment with Sunphenon (50 μg/mL)/ Polyphenon 60 (50 μg/mL) significantly (p<0.05) down-regulated it when compared to the untreated control group. Subsequent analysis of DNA degeneration and caspase activation revealed that Sunphenon (50 μg/mL)/Polyphenon 60 (50 μg/mL) inhibited activation of caspase-3 and showed an inhibitory effect on DNA degradation. From this result, we know that, in stress conditions, μ-calpain may be involved in the muscle atrophy through the suppression of myogenin and MyoD. Moreover, Sunphenon may regulate the skeletal muscle genes/promote skeletal muscle recovery by the up-regulation of myogenin and MyoD and suppression of μ-calpain and inflammatory pathways and may regulate the apoptosis pathways. Our findings suggest that dietary supplementation of Sunphenon might reduce inflammatory events in muscle-associated diseases, such as myotube atrophy.

Muscular expression of myogenin in V-pattern exotropia with inferior oblique overaction patients / 国际眼科杂志(Guoji Yanke Zazhi)

Abstract?AlM: To study the number of myogenin - positive activated satellite cells in the inferior oblique muscles and the medial muscles of V- pattern exotropia with inferior oblique overaction, to further explore the possible etiological factors of V-pattern exotropia with inferior oblique overaction.? METHODS: The inferior oblique muscles and the medial muscles were cut from V - pattern exotropia patients with inferior oblique overaction during strabismus operation treated as the strabismus group. Cross sections were stained immunohistochemically for the presence of activated satellite cells, as identified by myogenin immunoreactivity. The inferior oblique muscles and the medial muscles were obtained from the corneal transplant donors ( six eyes of six cases) , which treated as the control group.? RESULTS: The frequency of myogenin - positive satellite cells of the inferior oblique muscles was (22. 7± 7.03)% and (4. 2±0. 75)% in the strabismus group and the control group. Significant differences existed in the expression of myogenin in two groups (P<0. 05). Again, the frequency of myogenin-positive satellite cells of the medial muscles was (2. 2±0. 75)% and (4. 5±1. 05)% in the strabismus group and the control group. Significant differences also existed in the expression of myogenin in two groups (P<0. 05).?CONCLUSlON: lt is first report that myogenin-positive satellite cells presents in extraocular muscles of V -pattern exotropia with inferior oblique overaction. The current results suggest that myogenin is one of possible etiological factors of V-pattern exotropia with inferior oblique overaction.

Os efeitos do laser de baixa potência no processo de reparo muscular após criolesão em ratos / The effects of low-level laser therapy on muscle healing process after cryolesion

O objetivo deste estudo foi verificar os efeitos da laserterapia de baixa potência no comprimento de onda ?=780nm entre diferentes períodos de tratamento 7, 14 e 21 dias e verificar a dose (10J/cm2 ou 50J/cm2) que promove melhor reparo muscular através das análises histopatológicas e imunohistoquímicas. Foram utilizados 54 ratos machos divididos em 3 grupos: GC: grupo controle (criolesão, sem tratamento); G10: criolesão do músculo tibial anterior (TA) e tratados com laser dose 10J/cm² e G50: criolesão do músculo TA e tratados com laser dose 50J/cm² que foram subdivididos em 3 subgrupos (n=6): 7, 14 e 21 dias de tratamento. Os achados histopatológicos revelaram maior organização das fibras musculares dos grupos tratados com laser 10J/cm² e 50J/cm² durante os períodos 7 e 14 dias em relação ao grupo controle; no período 21 dias os grupos apresentaram semelhanças na reparação tecidual. Em relação à área da lesão os grupos tratados com laser 10J/cm² e 50J/cm² durante 7 dias obtiveram diminuição significativa (p ? 0.05) da área da lesão em relação ao grupo controle, sendo que os grupos 14 e 21 dias não apresentaram diferenças significativas entre eles. Na contagem dos vasos o grupo tratado com laser 10J/cm² no 14° dia apresentou aumento dos vasos em relação ao grupo tratado com dose 50J/cm², mas não em relação ao grupo controle. Nos tempos de 7 e 21 dias os grupos não apresentaram diferença significativa entre si. Com relação às análises imunohistoquímicas da myoD no período de 7 dias os grupos tratados com laser 10J/cm² e 50J/cm² apresentaram maior imunomarcação comparada com o grupo controle, no período 14 e 21 dias a imunomarcação estava ausente.

The objective of this study was to assess the effects of 780nm low-level laser therapy at different periods of 7, 14 and 21 days after cryolesion, including the dose (10 or 50J/cm2) to promote a better muscle repair evidenced by histopathological and immumohistochemical analyses. Fifty-four male rats were divided into three groups: injured control group (CG) - injured animals without any treatment; injured 780nm laser treated group, at 10 J/cm² (G10) and injured 780nm laser treated group, at 50 J/cm² (G50). Each group was divided into 3 subgroups (n=6): 7, 14 and 21 days post-injury. Histopathological findings revealed better-organized muscle fibers in the G10 and G50 during the periods of 7 and 14 days compared to CG. The G10 and G50 during 7 days showed a significant reduction (p? 0.05) of lesion area compared to CG, without differences between groups treated for 14 and 21 days. The G10 showed an increase of the amount of vessels after 14 days compared to the G50, but not in relation to controls. With regard to the immumohistochemical analyses of the MyoD factor, The G10 and G50 during 7 days showed higher concentrations of immunomarkers than controls.

Changes of myogenin expression in long-term denervated human posterior cricoarytenoid muscles / 第二军医大学学报

Objective: To investigate the change in myogenin expression at different time in long-term denervated human posterior cricoarytenoid muscles (PCAMs), so as to provide a theoretical basis for timing of reinnervation. Methods: Thirty-eight specimens of denervated human PCAMs were divided into 4 groups according to the period of denervation: 6-12 months denervation group(n= 12), 1-2 years denervation group (n = 10), 2-3 years denervation group (n = 8), and over 3 years denervation group(n=8). Another 12 specimens of normal PCAMs served as control. The patients in all groups were age- and sex-matched. The expression of myogenin protein and mRNA was studied using immunofluorescence staining and real-time PCR analysis, respectively. Results: Immunofluorescence staining showed that the positive myogenin expression was mainly found in the myonuclei of PCAMs with a denervation period less than 3 years; no positive staining was found in the myonuclei of control group. The expression of myogenin in myonuclei and the ratio of positive cells were up-regulated in the 6-12 month denervation group compared with those in the control group; the expression and the ratio peaked in 1-2 years denervation group and decreased again in the 2-3 years denervation group, but was still significantly higher than those of the control group (all P< 0.01). There was hardly any expression of myogenin 3 years after denervation. Results of RT-PCR showed no myogenin mRNA expression in the control group; the expression in 1-2 years, 2-3 years, and more than 3 years denervation groups were 4 times, 64 times, and half that of the 6-12 months denervation group, respectively (all P<0.05). Conclusion: It is indicated that there is a potential for muscle regeneration within 3 years of denervation.

Changes of myogenin expression in long-term denervated human posterior cricoarytenoid muscles / 第二军医大学学报

Objective:To investigate the change in myogenin expression at different time in long-term denervated human posterior cricoarytenoid muscles(PCAMs),so as to provide a theoretical basis for timing of reinnervation.Methods: Thisty-eight specimens of denervated human PCAMs were divided into 4 groups according to the period of denervation: 6-12 months denervation group(n=12),1-2 years denervation group(n=10),2-3 years denervation group(n=8),and over 3 years denervation group(n=8).Another 12 specimens of normal PCAMs served as control.The patients in all groups were age-and sex-matched.The expression of myogenin protein and mRNA was studied using immunofluorescence staining and real-time PCR analysis,respectively.Results: Immunofluorescence staining showed that the positive myogenin expression was mainly!found in the myonuclei of PCAMs with a denervation period less than 3 years;no positive staining was found in the myonuclei of control group.The expression of myogenin in myonuclei and the ratio of positive cells were up-regulated in the 6-12 month denervation group compared with those in the control group;the expression and the ratio peaked in 1-2 years denervation group and decreased again in the 2-3 years denervation group,but was still significantly higher than those of the control group(all P