[Effects of Janus kinase/signal transduction and activator of transcription 3 pathway inhibitor in skeletal muscle function in severely burned rats and its mechanism].

Objective: To observe the functional changes of skeletal muscle in severely burned rats, and to investigate the effects and possible mechanisms of Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) pathway inhibitor in skeletal muscle function. Methods: The experiment research method was applied. One hundred and twenty male Wistar rats of 8-week-old were divided into sham injury group, simple burn group, and burn+JAK/STAT3 inhibitor group according to the random number table, with 40 rats in each group. Rats in simple burn group and burn+JAK/STAT3 inhibitor group were inflicted with 50% total body surface area full-thickness scald on the back and abdomen, and rats in sham injury group were sham injured. Rats in burn+JAK/STAT3 inhibitor group were intraperitoneally injected with JAK/STAT3 inhibitor ruxolitinib. On post injury day (PID) 0 (immediately), 1, 4, 7, and 14, 8 rats in each group were used to measure the specific force generated by extensor digitorum longus in optimal length stimulated with pulse frequency of 20, 40, 60, 80, 100, 120, 140, and 160 Hz using a multichannel electrophysiological instrument, and specific force in fatigue period of extensor digitorum longus in optimal length stimulated with pulse frequency of 50 Hz for 0, 10, 20, 30, 60, 120, 180, 240, and 300 s. On PID 0, 1, 4, 7, and 14, carbonyl compound content of extensor digitorum longus was determined by ultraviolet spectrophotometry, and ATP content of extensor digitorum longus was determined by micrometry. Data were statistically analyzed with analysis of variance for repeated measurement, analysis of variance for factorial design, Bonferroni method, and t test. Results: Compared with those of sham injury group, specific forces of extensor digitorum longus of rats in simple burn group were significantly decreased after being stimulated with all the pulse frequency on PID 0, 1, 7, and all the pulse frequency except for 20 Hz on PID 4, and pulse frequency of 20 and 40 Hz on PID 14 (P<0.05 or P<0.01). Compared with those of simple burn group, specific forces of extensor digitorum longus of rats in burn+JAK/STAT3 inhibitor group were significantly increased after being stimulated with all the pulse frequency except for 20 Hz on PID 1 and all the pulse frequency on PID 4, 7, and 14 (P<0.05 or P<0.01). Compared with those of sham injury group, specific forces of extensor digitorum longus of rats in simple burn group were significantly decreased in fatigue period at all the time points post injury and stimulation time points except for 240 s on PID 7 (P<0.05 or P<0.01). Compared with those of simple burn group, specific forces of extensor digitorum longus of rats in burn+JAK/STAT3 inhibitor group were obviously increased in fatigue period at all the stimulation time points except for 60 and 300 s on PID 1 and 240 s on PID 4, and all the stimulation time points on PID 7 and 14 (P<0.05 or P<0.01). The carbonyl compound content of extensor digitorum longus of rats in simple burn group on PID 0, 1, 4, 7, and 14 was (0.651±0.155), (0.739±0.194), (0.618±0.086), (0.813±0.162), (0.615±0.115) nmol/mg, which were obviously higher than (0.196±0.019), (0.156±0.004), (0.169±0.023) (0.156±0.027), (0.175±0.008) nmol/mg in sham injury group (t=7.219, 6.491, 10.938, 9.182, 11.589, P<0.01) and (0.538±0.069), (0.369±0.059), (0.273±0.061), (0.334±0.109), (0.318±0.101) nmol/mg in burn+JAK/STAT3 inhibitor group (t=2.446, 4.689, 8.355, 5.754, 6.097, P<0.05 or P<0.01). The ATP content in extensor digitorum longus of rats in simple burn group on PID 1, 4, 7, and 14 was obviously lower than that in sham injury group (t=7.159, 7.591, 7.473, 4.026, P<0.01) and burn+JAK/STAT3 inhibitor group (t=2.295, 2.575, 2.453, 2.997, P<0.05). Conclusions: After severe burn, the specific force of extensor digitorum longus in rats decreased significantly after being stimulated with different pulse frequencies, and the extensor digitorum longus in rats was prone to fatigue. Blocking the JAK/STAT3 signaling pathway can reduce the oxidative stress of muscle protein and increase ATP content, thereby reducing the muscle strength decline caused by burn injury and improving the muscle strength decline during fatigue period.

Numerical modelling to assess the tear force of human capsulotomy margin.

Tear force of the capsulotomy edge is a paramount parameter to affect the surgical safety in cataract surgery. This paper aimed to investigate the stretch force of the capsulotomy edge using finite element (FE) method. A FE model of capsule bag was developed to simulate dynamic response of the lens capsule to the stretch of retractors. The failure criterion based on the distortion energy theory was applied to predict the rupture of the anterior capsule. The simulation results showed a good agreement with the experimental data reported in the literature. Sensitivity studies were then conducted to evaluate the effect of the various parameters on tear force, including the stretching velocity, capsulorhexis dimension, age, retractor width and shape, and rim morphology. The rupture force was proportional to the stretching velocity, capsulorhexis dimension and retractor width, while the age showed the opposite trend. In addition, the retractor shape has a greatly effect on the tear force and the rim of continuous curvilinear capsulorhexis (CCC) has the higher tear resistance. This work can contribute to the understanding of the regularity for capsule rupture.

Influence of needling conditions on the corneal insertion force.

Needle insertion plays an important part in the process of corneal graft surgery. In this paper, a three-dimensional symmetry model of the human cornea is constructed using the finite element method. Simplification of specific optic physiology is defined for the model: The cornea constrained by the sclera is presented as two layers consisting of epithelium and stroma. A failure criterion based on the distortion energy theory has been proposed to predict the insertion process of the needle. The simulation results show a good agreement with the experimental data reported in the literature. The influence of needling conditions (e.g. insertion velocity, rotation parameters and vibration parameters) on the insertion force are then discussed. In addition, a multi-objective optimization based on particle swarm optimization (PSO) is applied to reduce the insertion force. The numerical results provide guidelines for selecting the motion parameters of the needle and a potential basis for further developments in robot-assisted surgery.

[Effects of insulin therapy on skeletal muscle wasting in severely scalded rats and its related mechanism].

Objective: To explore the effects of insulin therapy on skeletal muscle wasting (SMW) in severely scalded rats and its related mechanism. Methods: Totally 48 male Wistar rats aged 7-8 weeks were divided into simple scald (SS) group and insulin therapy (IT) group according to the random number table, with 24 rats in each group. After weighing the body mass and measuring the blood glycemic level of the tail end with a glucometer, the rats in the two groups were immersed in hot water at 94 ℃ for 12 seconds to make a full-thickness dorsal scald model involving 30% total body surface area. Rats in group IT were subcutaneously injected with 1 U/kg insulin glargine at 8: 00 a day from post injury day (PID) 1 to 7, whilst rats in group SS were given the same amount of normal saline. Rats in the two groups were given 10 mL/kg enteral nutritional emulsion by intragastric infusion at 8: 00 (after insulin administration), 13: 00, and 18: 00 a day respectively from PID 1 to 7. The blood glycemic levels of tail end of rats in the two groups were measured by glucometer before insulin administration on PID 1-4, 6, and 7 and on every morning of PID 8, 9, 11, 12, and 14. The body mass of rats in the two groups on PID 14 without any treatment was weighed. Eight rats from each group were collected respectively on PID 4, 7, and 14 to harvest tibialis anterior muscle (TAM) samples. The mass of TAM on PID 14 was weighed. The ultrastructural changes of TAM myocytes on PID 7 were observed with transmission electron microscope. The apoptotic rates of TAM myocytes on PID 4, 7, and 14 were assessed by the assay of terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphate-biotin nick end labeling, the expressions of cysteine-aspartic protease-3 (caspase-3) of TAM on PID 4, 7, and 14 were detected with immunohistochemistry, and protein expressions of endoplasmic reticulum (ER) stress (ERS) associated proteins glucose-regulated protein 78 (GRP78), CCAAT/enhancer binding protein-homologous protein (CHOP), and activated caspase-12 of TAM on PID 4, 7, and 14 were detected with Western blotting. Data were processed with completely random design t test, analysis of variance for repeated measurement, analysis of variance for factorial design, t test, and Bonferroni correction. Results: The blood glycemic level and body mass of rats in the two groups before injury were similar (t=0.204, 0.405, P>0.05). There were no statistically significant differences in blood glycemic levels of rats between the two groups on PID 1, 6, 9, 11, 12, and 14 (t=0.229, 3.339, 1.610, 0.178, 0.181, 0.079, P>0.05). Compared with those of group SS, blood glycemic levels of rats in group IT were significantly lower on PID 2, 3, 4, 7, and 8 (t=7.245, 4.165, 4.609, 4.018, 3.995, P<0.05 or P<0.01). On PID 14, the body mass and TAM mass of rats in group IT were (271±19) g and (0.47±0.05) g respectively, both obviously higher than (254±12) g and (0.43±0.04) g of group SS (t=2.159, 2.375, P<0.05). On PID 7, nuclear pyknosis and deformation, chromosome misdistribution, and ER swelling in TAM myocytes of rats in group SS were observed; the apoptotic alterations and ER swelling of TAM myocytes were alleviated in rats of group IT as compared with those of group SS. The apoptotic rates of TAM myocytes of rats in group IT were obviously lower than those of group SS on PID 4, 7, and 14 (t=4.262, 9.153, 9.799, P<0.01). The expressions of caspase-3 in TAM of rats in group IT were obviously lower than those of group SS on PID 7 and 14 (t=10.429, 7.617, P<0.01). Compared with those of group SS, the protein expressions of GRP78 were obviously increased on PID 4 and 14 (t=4.172, 4.437, P<0.05), the protein expressions of activated caspase-12 were obviously decreased on PID 7 and 14 (t=11.049, 11.181, P<0.01), and the protein expressions of CHOP were obviously decreased on PID 4, 7, and 14 (t=13.837, 9.572, 6.930, P<0.01) in TAM of rats in group IT. Conclusions: Insulin therapy may reduce skeletal muscle myocytes apoptosis and SMW by alleviating ERS in rats with severe scald.

[SIRT1 signaling pathway mediated the protective effects on myocardium of rats after endurance training and acute exhaustive exercise].

Objective: To detect the expression of SIRT1 and Ac-FOXO1 in rats after endurance training and acute exhaustive exercise, and explicit the myocardial protective effect of SIRT1. Methods: Rats were randomly divided into four groups: control group(n=20), exhaustive exercise group (E group, n=20), exhaustive exercise group + endurance training (TE group, n=18), exhaustive exercise group + endurance training + selective SIRT1 inhibitor (TSE group, n=17). The Control and E groups were fed routinely for 5 weeks. The TE and TSE groups were subjected to swimming exercise for 5 weeks for endurance exercising. The TSE group was intraperitoneally injected with selective SIRT1 inhibitor Sirtinol(2 mg/kg) at 30 minutes before endurance exercising. The E, TE and TSE groups were subjected to exhaustive exercise. The myocardial tissues of rats were collected after exhaustive exercise. Real-time polymerase chain reaction (PCR) and Western blot analysis were performed to detect the myocardial mRNA and protein expressions of SIRT1 and Ac-FOXO1. The myocardial protein expression of Bax and Bcl-2 was also detected by Western blot. Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay was used to assess the apoptosis of myocardial cells. Results: Compared with Control group, the SIRT1 and Bcl-2 expression in the myocardial tissue was obviously decreased, while the Ac-FOXO1, Bax, and the myocardial cell apoptosis were significantly increased in E group (all P<0.01). Compared with E group, the expression of SIRT1 and Bcl-2 was obviously up-regulated (both P<0.01), while the Ac-FOXO, Bax and the myocardial cell apoptosis was significantly reduced in TE group (all P<0.01). Compared with TE group, the SIRT1 and Bcl-2 expression was obviously lower (both P<0.01), while Ac-FOXO1, Bax, and the cell apoptosis were significantly higher in group TSE (all P<0.01). Conclusion: Endurance training could protect myocardium by reducing the myocardial oxidative stress injury and apoptosis via activating SIRT1 signaling pathway, up-regulating the myocardial expression of SIRT1 and regulating the deacetylation of FOXO1.

[Effects of leucine on adipogenesis in 3T3-L1 preadipocytes during and after differentiation].

OBJECTIVE: To observe the effects of leucine on adipogenesis in 3T3-L1 preadipocyte during and after differentiation, and to investigate possible mechanisms. METHODS: Respectively, 0.0 (control), 0.5, 1.0 and 2.0 mmol/L leucine was added in 3T3-L1 cells and cell proliferation was measured by MTT. Then, 3T3-L1 preadipocyte was induced to differentiate. Leucine was added during whole differentiation period, or after differentiation for 4 days. The cells were stained with Oil Red O dye to observe lipid droplet. The culture media were collected and used to determine glycerol contents. Meanwhile, protein expressions related to lypolytic enzymes, leptin signaling pathway were determined by Western blot. RESULTS: MTT result showed that cell viabilities were (100.00±12.10)%, (102.73±12.38)%, (103.94±14.65)%, (108.70±5.05)% in 0.0, 0.5, 1.0 and 2.0 mmol/L leucine groups, respectively, there were no significant differences in cell proliferation among 4 groups (F=1.07, P=0.383). When 0.0, 0.5, 1.0 and 2.0 mmol/L leucine was added during differentiation, the relative number of lipid droplet was 1.00±0.06, 0.94±0.09, 0.82±0.08 and 0.79±0.04, respectively (F=11.74, P<0.001), and it was significantly lower in 1.0 and 2.0 mmol/L leucine groups than in control group (P=0.002 and P<0.001, respectively). There was no significant difference in lipid droplet when leucine was added after differentiation (F=0.16, P=0.924). When leucine was added during differentiation, the increment of glyceride contents in medium was (65.04 ± 11.75), (71.45 ± 23.71), (79.37 ± 17.63) and (110.32 ± 25.36) µmol/L, respectively (F=2.92, P=0.100). And it was significantly higher in 2.0 mmol/L leucine group (110.32 ± 25.36) µmol/L than in control group (65.04 ± 11.75) µmol/L (t=2.73, P=0.026). No significant difference of the increment of glyceride contents among 4 groups was observed when leucine was added after differentiation (F=0.80, P=0.528). Western blot results showed that leucine treatment during differentiation upregulated expression level of hormone-sensitive lipase phosphorylation (after 0.0 and 2.0 mmol/L leucine treatment,the protein levels were 1.00 ± 0.08 vs. 2.54 ± 0.27, P<0.001) , and downregulated the protein expression levels of perilipin A, leptin and leptin-related pathway, such as leptin receptor, Janus kinase 2 and suppressor of cytokine signaling-3 (after 0.0 and 2.0 mmol/L leucine was added, the protein levels were (1.00 ± 0.03) vs. (0.31 ± 0.07) , (1.00 ± 0.08) vs. (0.22±0.07) , (1.00±0.07) vs. (0.21 ± 0.04) , (1.00 ± 0.03) vs. (0.35 ± 0.05) , (1.00 ± 0.06) vs. (0.34 ± 0.05) , P<0.001). Leucine treatment after differentiation had no effects on these protein expressions (all P>0.05). CONCLUSION: Leucine inhibits adipogenesis during 3T3-L1 preadipocyte differentiation by the regulation of lypolytic enzymes and leptin signaling pathway; however, leucine has no effect on adipogenesis when differentiation completed.