Atrophic age-related macular degeneration (AMD) does not show obvious loss of visual function in the early stage, so it is not easy to be taken seriously. In the advanced stage, most of the patients suffered from macular area retinal map atrophy, which affected night vision and central vision. Drugs currently used in clinical or clinical trials to treat atrophic AMD include drugs for improving choroidal perfusion, reducing the accumulation of harmful substances, preventing oxidative stress injury, inhibiting inflammatory reactions, as well as neuroprotectants and lipid metabolism drugs. Stem cell transplantation for atrophic AMD is currently the most promising treatment. In theory, it is feasible to replace atrophic AMD with retinal photoreceptor cells and RPE cells derived from human stem cell differentiation. However, there are still many problems to be solved, such as how to improve the efficiency of directional differentiation of seed cells and how to ensure the safe and effective RPE cell transplantation and survival after transplantation. At present, several studies have found that multiple locus mutations are associated with atrophic AMD, so gene therapy also plays an important role in the development of the disease.
Objective To investigate the effect of exogenous erythropoietin (EPO) on the denervated muscle atrophy. Methods Twenty-four SD male rats, weighting 200-220 g were made the models of denervated gastrocnemius muscle after sciatic nerves were transected under the piriform muscle at the right lower leg, and were randomly divided into two groups (n=12). rhEPO (2 500 U/kg) was injected daily into the denervated gastrocnemius muscle in EPO group, and normal sal ine was injected into the denervated gastrocnemius muscle in control group. To observe the general state of health of the experimental animal, the muscle wet weight, the muscle cell diameter, the cross section area, the protein amount, thepercentage of the apoptotic muscle cells, and the Na+-K+-ATPase and Ca2+-ATPase activities were measured 2 and 4 weeks after operation. Results All experimental animals were survived during experiment without cut infection, and all animals could walk with pull ing the right knee. At 4 weeks after operation, 7 cases showed ulcer in the right heel, inculding 5 in the control group and 2 in the EPO group. At 2 and 4 weeks after operation, the muscle wet weight in EPO group was (885.59 ± 112.35) and (697.62 ± 94.74) g, respectively; in control group, it was (760.63 ± 109.05) and (458.71 ± 58.76) g, respectively; indicating significant differences between two groups (P lt; 0.01). The protein amount in EPO group was (77.37 ± 5.24) and (66.37 ± 4.87) mg/mL, respectivly；in control group, it was (65.39 ± 4.97) and (54.62 ± 6.32) mg/mL；indicating significant differences between two groups (P lt; 0.01). At 2 and 4 weeks after operation, the myofibrillar shapes were nearly normal in EPO group while there were muscle fiber atrophy, some collapse and obviously hyperblastosis between muscle bundle. There were significant differences in the muscle cell diameter and the cross section between two groups (P lt; 0.01). However, the percentage of the apoptotic muscle cells was 11.80% ± 1.74% and 28.47% ± 1.81% in control group, respectively, which was significantly smaller than that in EPO group (21.48% ± 2.21% and 55.89% ± 2.88%, P lt; 0.01). At 2 and 4 weeks after operation, Na+-K+-ATPaseand Ca2+-ATPase activities in EPO group were higher than those in control group (P lt; 0.01). Conclusion EPO can delay the denervated muscle atrophy.
ObjectiveTo observe the peripapillary atrophy (PPA) and peripapillary choroidal vascularity index (CVI) in patients with different degrees of myopia and to analyze their correlations. MethodsA cross-sectional clinical study. From September 2021 to December 2021, 281 mypoic patients of 281 eyes treated in Eye Hospital of Wenzhou Medical University at Hangzhou were included in this study, and the right eye was used as the treated eye. There were 135 eyes in 135 males and 146 eyes in 146 females. The age was 28.18±5.78 years. The spherical equivalent refraction (SE) was －5.13±2.33 D. The patients were divided into three groups: low myopia group (group A, －3.00 D ＜SE≤－0.50 D), moderate myopia group (group B, －6.00 D≤SE≤－3.00 D);high myopia group (group C, SE＜－6.00 D). The spherical equivalent refraction was statistically different among the three groups (H=241.353, P＜0.05). All of the affected eyes were examined by swept-source optical coherence tomography. Combined with B-scan image,assessment and area measurement of β area, γ area (β-PPA and γ-PPA) were carried out on the en-face image. After binarization of the collected images, the nasal, superior, temporal and inferior CVI of the optic disc were calculated. For comparison between groups, one-way ANOVA was used for continuous variables with normal distribution, Kruskal-Wallis test was used for continuous variables with abnormal distribution, and categorical variables were used χ2 inspection. Linear regression analysis was used for the relationship between β-PPA and γ-PPA area and peripapillary CVI of different regions. Linear regression analysis was used to evaluate the relationships between the area of peripapillary atrophy and peripapillary choroidal vascularity index in different regions. ResultsThere was no statistical difference in the incidence of β-PPA among the three groups (χ2=4.672, P=0.097). The incidence of γ-PPA in group A was lower than that in group B anc C, and the difference was statistically different (χ2=33.053, P＜0.001), in which both group A was lower than group B and C. Among the three groups, the area of β-PPA and γ-PPA was statistically significant (H=36.535, 39.503; P＜0.001, 0.001); the β-PPA area of group A and B was lower than that of group C; the γ-PPA area was group A＜group B＜group C. Peripapillary CVI of different regions in group A, group B and group C was statistically significant (F=11.450, 5.037, 6.018, 4.489; P＜0.05). The temporal CVI in group C was lower than that in group A and B; The inferior CVI of group C was lower than that of group A, and the superior and nasal CVI of group B and C were lower than that of group A. In multivariate analysis, SE (β=0.374, P＜0.001), temporal CVI (β=－0.299, P＜0.001) were correlated with the area of β-PPA (adjusted R2=296, P＜0.001); AL (β=0.452, P＜0.001), temporal CVI (β=－0.220, P＜0.001) were correlated with the area of γ-PPA (adjusted R2=0.309, P＜0.001). ConclusionsThe incidence and area of γ-PPA are increased in the higher degree of myopia group. The area of γ-PPA is positively correlated with the axial length, and both the area of β-PPA and γ-PPA are negatively correlated with temporal CVI.
Objective To investigate the role of cysteinyl aspartate specific proteinase-3 (Caspase-3)/ gasdermin-E (GSDME)-mediated pyroptosis in skeletal muscle atrophy induced by cigarette smoke in mice.Methods To construct a mouse model of COPD, C57BL/6 mice were exposed to cigarette smoke (CS) for 24 weeks. HE staining was used to observe the changes in the morphology of the gastrocnemius muscle in mice. Immunohistochemistry was used to detect the expression of pyroptosis-related proteins in gastrocnemius muscle. To construct a model of skeletal muscle cell atrophy in vitro, C2C12 myoblasts were induced to differentiate into skeletal muscle cells with 2% horse serum, and then skeletal muscle cells were treated with cigarette smoke extract (CSE). Skeletal muscle cells were further treated with the caspase-3 inhibitor Z-DEVD-FMK and the GSDME inhibitor Dimethyl fumarate (DMF) to explore the effects of inhibition of caspase-3/GSDME on CSE-induced skeletal muscle cell atrophy. To observe the effects of TNF-α on the expression of caspase-3 and GSDME proteins as well as the impact on myotubes, skeletal muscle cells were stimulated with tumor necrosis factor-alpha (TNF-α). Western blotting was applied to detect protein expression levels of caspase-3 and GSDME in skeletal muscle cells. Hoechst 33342/ Hoechst33342/ Propidium Iodide (PI) staining was applied to detect the PI-positive rate of skeletal muscle cells. The lactate dehydrogenase (LDH) release of C2C12 myotubes was measured by LDH release test. Immunofluorescence was used to detect changes in myotube diameter. Results CS-induced skeletal muscle atrophy was observed in mice, accompanied by increased pyroptosis- associated proteins (c-caspase-3 and GSDME-N) (P<0.05). CSE also induced elevated c-caspase-3 and GSDME-N expression in C2C12 cells , resulting in increased LDH release, positive ratio of PI, along with reduced myotube diameter (P<0.05). In addition, TNF-α promotes myotube atrophy and the expression of cleaved-caspase-3 and GSDME-N proteins in skeletal muscle cells. ConclusionCS can induce skeletal muscle atrophy through activated TNF-α/Caspase-3/GSDME-mediated pyroptosis.
Objective To observe the delaying effect of neural stem cell (NSC) transplantation on denervated muscle atrophy after peri pheral nerve injury, and to investigate its mechanism. Methods NSCs were separated from the spinal cords of green fluorescent protein (GFP) transgenic rats aged 12-14 days mechanically and were cultured and induced to differentiate in vitro. Thirty-two F344 rats, aged 2 months and weighed (180 ± 20) g, were randomized into two groups (n=16 per group). The animal models of denervated musculus triceps surae were establ ished by transecting right tibial nerve and commom peroneal nerve 1.5 cm above the knee joints. In the experimental and the control group, 5 μL of GFP-NSCsuspension and 5 μL of culture supernatant were injected into the distal stump of the tibial nerve, respectivel. The generalcondition of rats after operation was observed. At 4 and 12 weeks postoperatively, the wet weight of right musculus tricepssurae was measured, the HE staining, the Mallory trichrome staining and the postsynaptic membrane staining were adopted for the histological observation. Meanwhile, the section area of gastrocnemius fiber and the area of postsynaptic membrane were detected by image analysis software and statistical analysis. Results The wounds in both groups of animals healed by first intension, no ulcer occurred in the right hind l imbs. At 4 and 12 weeks postoperatively, the wet weight of right musculus triceps surae was (0.849 ± 0.064) g and (0.596 ± 0.047) g in the experimental group, respectively, and was (0.651 ± 0.040) g and (0.298 ± 0.016) g in the control group, respectively, showing a significant difference (P lt; 0.05). The fiber section area of the gastrocnemius was 72.55% ± 8.12% and 58.96% ± 6.07% in the experimental group, respectively, and was 50.23% ± 4.76% and 33.63% ± 4.41% in the control group, respectively. There were significant differences between them (P lt; 0.05). Mallory trichrome staining of muscle notified that there was more collagen fiber hyperplasia of denervated gastrocnemius in the control group than that in the experimental group at 4 and 12 weeks postoperatively. After 12 weeks of operation, the area of postsynaptic membrane in the experimental group was (137.29 ± 29.14) μm2, which doubled that in the control group as (61.03 ± 11.38) μm2 and was closer to that in normal postsynaptic membrane as (198.63 ± 23.11) μm2, showing significant differences (P lt; 0.05). Conclusion The transplantation in vivo of allogenic embryonic spinal cord NSCs is capable of delaying denervated muscle atrophy and maintaining the normal appearance of postsynaptic membrane, providing a new approach to prevent and treat the denervated muscle atrophy cl inically.
Objective To investigate the delay of the denervated skeletal muscle atrophy with the method of restraining the increment of the connective tissues by tetrandrine and hormone. Methods The left hind limbs of 42 male adult SD rats were made into models of the denervated gastrocnemius, and then the rats were randomly divided into 3 groups, with 14 rats in each. In Group A, tetrandrine (8 mg/L）was injected into the denervated gastrocnemius; in Group B, triamcinolone acetonide（1.6 g/L） was injected; in Group C (the control group),normal saline was injected. Enough samples were obtained according to the different observation indexes at 30 days after operation. Electromyography, muscle wet weight measurement, light microscopy,electron microscopy,and microimage analysis were performed. ResultsThe fibrillation potential amplitude was 0.195 8±0.041 9 μV in Group A and 0.185 2±0.050 3 μV in Group B, and there was no significant difference betweenthe two groups (Pgt;0.05). However,in Group C the fibrillation potential amplitude was 0.137 7±0.058 9μV. The fibrillation potential amplitude was significantly greater in Group A than in Group C（Plt;0.05）. The muscle wet weight was 1.740 0±0.415 9 g in Group A and 1.940 1±0.389 4 gin Group B, and there was no significant difference between the two groups(Pgt;0.05).However, in Group C the muscle wet weight was 0.800 0±0.100 0 g. The muscle wet weight was significantly greater in Group A than in Group C（Plt;0.05）.The microscopy showed that more remarkable atrophy occurred in the control group. The muscle fibers were more complete, thicker and larger, with more nuclei and clearer cross-lines. More connective tissue and flat cells could be observed in Groups A and B. The myogenic protein amount was 440.124 2±46.135 6 in Group A and 476.211 4±41.668 8in Group B, and there was no significant difference between the two groups(Pgt;0.05).However, in Group C the amount was 380.040 0±86.315 9.The myogenic protein amount was significantly greater in Group A thanin Group C（Plt;0.05）. The muscle fiber number, diameter, cross section, and connective tissue increment were all significantly greater in Group A than in Group C（Plt;0.05）; however, there wasno significant difference between Groups A and B (Pgt;0.05). The electron microscopy showed that there were more degeneration changes, such as muscle silk disorder, chondriosome disappearance, and hepatin reduction, could be observed inGroup C than in Groups A and B. Conclusion Tetrandrine and hormone can delay the denervated skeletal muscle atrophy by restraining the increment of the connective tissues.
The present paper intends to discuss the antioxidant and tyrosinase inhibition effect of solanesol from three aspects of ultraviolet radiation and free radical scavenging. The paper makes a survey on diurnal variation rule of the minimum ultraviolet transmittance and ultraviolet transmittance of solanesol, hydroxyl (·OH) free radical scavenging method of Smirnoff reaction system model, superoxide anion O2-· free radical scavenging method of pyrogallol autoxidation, and the inhibitory effect of solanesol on tyrosinase activity by enzyme kinetic method. The experiment results showed that solanesol could effectively scavenge lipid radicals to block lipid peroxidation, and inhibit effects on tyrosinase. Solanesol is a natural extract which could be used to prevent senile atrophy of human skin and senile plaque.
Leber hereditary optic neuropathy is an optic neuropathy associated with mitochondrial DNA. The disease affects young men mainly, which is considered to be due to denaturation of the retinal nerve ganglion cell and axonal loss of optic nerve, leading to optic atrophy. Nowadays, there are some development in studying Leber hereditary optic neuropathy by optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA). It is great help to know the disease, forecast the progression of disease, and take action on intervention. In addition, there is a lack of in-depth study on OCT and OCTA characteristics among different mutation sites of LHON, different genders of the same site, different families of the same site or even different branches of the same family. It is expected to be improved in the future work.
Objective To find the new mutations of Leber's hereditary optic neuropathy (LHON). Methods Two LHON families were enrolled in this study. The probands and all maternal members in this two families were underwent ophthalmologic examinations. The ages of probands were seven and 14 years old respectively. A total of 358 healthy adults were enrolled in this study as control group. The genomic DNA from whole blood of participants were extracted. The entire mitochondrial genome of probands were PCR amplified and sequenced in 24 overlapping fragments using primers as designed. At the same time, the mtDNA of maternal relatives and 358 controls were also detected. Fourteen primate species were selected from GenBank to analyzed the phylogenetics of mitochondrial sequence. Results There was no ND4 G11778A, ND1 G3460A, ND6 T14484C mutational site in all maternal members. Molecular analysis of mtDNA in this two families identified the homoplasmic tRNAGluA14683G mutation and distinct set of variants belonging to the Asian haplogroup F1a1 and G2. The site was at theTpsi;C stem oftRNAGlu and extremely conserved among 14 primate species. It was anticipated that the A14683G increased the highly conserved C-G basepairing. Furthermore, the A14683G was absence in control group. Conclusion The tRNAGluA14683G mutation is likely a new mutation associated with LHON.
ObjectiveThe aim was to summarize the seizure and video electroencephalogram (VEEG) characteristics of Dyke-Davidoff-Masson syndrome (DDMS). Methods The case data of four patients with Dyke-Davidoff-Masson syndrome (DDMS) who attended the Epilepsy Center of Hunan Provincial Brain Hospital from March 2022 to March 2023 were retrospectively analyzed to summarize the clinical manifestations of their seizures and the characteristics of their video electroencephalogram (VEEG). Results One case of symptomatic epilepsy with focal seizures; VEEG showed poor background activity alpha rhythmic modulation, amplitude modulation, and increased distribution of slow wave activity in the left frontal and temporal regions; bilateral frontal-central and anterior-temporal regions (more so on the left side), with sharp and slow composite wave issuance.Two cases of symptomatic epilepsy with focal seizures progressing to generalized seizures; in one case, the VEEG showed: background activity α-rhythmic modulation, amplitude modulation is possible, the left frontal, central, anterior temporal region slow wave increase; the left frontal central, parietal anterior temporal region spike-like slow wave activity mixed with spike wave, spike-slow complex wave short-medium-range issuance; the other VEEG showed: background activity α-rhythmic modulation, amplitude modulation is possible, the right frontal central, anterior temporal region slow wave increase; right frontal, central, and anterior temporal region for the famous medium-extremely high-high-amplitude slow wave activity mixed with spike wave, spike-slow complex wave short-medium-range issuance. One case of symptomatic epilepsy with generalized seizures; VEEG showed bilateral occipital alpha rhythm asymmetry, right occipital region <50% of the left side, poor regulation and amplitude modulation; bilateral frontal pole, frontal region, anterior temporal region spike and spiking slow complex wave discharges (right side was prominent), and right pterionic electrodes, anterior temporal and mesial temporal spike and spiking slow wave discharges. Conclusions Epileptic seizures are one of the main clinical manifestations of DDMS and most of them are consulted after a seizure, and their seizure types tend to be focal seizures or progress to generalized seizures, and most of them are drug-refractory epilepsies. The results of VEEG monitoring tend to be characterized by abnormal background activity, increased slow-wave activity, and the site of epileptogenic wave-like discharges tends to be in line with the site of cerebral softening foci or the site of the atrophic side of the brain as shown by cranial MRI.