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Objective@#To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro.@*Methods@#The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference t test, and Bonferroni correction.@*Results@#(1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t3 h=4.891, 5.890, 4.928; t6 h=9.790, 6.750, 10.590; t9 h=6.948, 6.764, 5.049, P<0.05 or P<0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 μmol/L capsaicin group was increased (t=10.488, P<0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t=4.372, 3.026, P>0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 μmol/L capsaicin group and hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (t=15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P<0.05 or P<0.01), which of hypoxia+ 10.0 μmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t=4.365, P<0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t=6.216, P<0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t=3.489, P<0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t=0.545, P>0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t=12.550, 7.442, P<0.01).@*Conclusions@#TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.
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Objective@#To analyze the clinical characteristics of early organ injury in elderly patients with severe burns and the effects on the prognosis of patients.@*Methods@#From January 2010 to August 2018, 62 patients with severe burns (43 men and 19 women, aged from 60 to 89 years at the time of admission) who were hospitalized in the Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the author′s affiliation), meeting the inclusion criteria, were included in elderly (E) group, and 124 patients with severe burns (86 men and 38 women, aged from 18 to 59 years at the time of admission) at the same term were included in young and middle-aged (YM) group. Treatment of patients in the 2 groups followed the conventional procedures of the author′s affiliation. The following data of patients in the 2 groups were retrospectively analyzed. (1) Fluid replacement volume and urine volume within the first and second post injury hour (PIH) 24 were recorded. The levels of hemoglobin, haematocrit, and blood lactic acid at admission, PIH 24 and 48 were recorded. (2) The creatine kinase isozyme-MB (CK-MB), total bilirubin, blood creatinine, oxygenation index, and blood platelet count at admission, at shock stage, and on post injury day (PID) 3 to 7 were collected. (3) The days of seriously or critically ill and deaths were recorded. Data were processed with chi-square test, group t test, Mann-Whitney U test, analysis of variance for repeated measurement, and Bonferroni correction.@*Results@#(1) There were no statistically significant differences in fluid replacement volume within the first and second PIH 24, and urine volume within the second PIH 24 between patients in the 2 groups (t=0.351, 1.307, 1.110, P>0.05). The urine volume of patients in group E within the first PIH 24 was significantly less than that in group YM (t=5.628, P<0.05). There were no statistically significant differences in levels of hemoglobin (t=0.011, 1.075, 0.239), haematocrit (t=0, 0.033, 0.199), and blood lactic acid (t=0.017, 1.002, 0.739) at admission, PIH 24 and 48 between patients in the 2 groups (P>0.05). (2) There were no statistically significant differences in levels of CK-MB at admission and on PID 3 to 7 between patients in the 2 groups (t=0.069, 0.001, P>0.05). The level of CK-MB of patients in group E at shock stage was significantly higher than that in group YM (t=4.017, P<0.05). There were no statistically significant differences in levels of total bilirubin at admission and on PID 3 to 7 between patients in the 2 groups (t=0.227, 0.002, P>0.05). However, the level of total bilirubin of patients in group E at shock stage was significantly higher than that in group YM (t=6.485, P<0.05). The levels of blood creatinine of patients in group E at admission and shock stage were significantly higher than those in group YM (t=4.226, 12.299, P<0.05 or P<0.01), while there was no statistically significant difference between them on PID 3 to 7 (t=0.693, P>0.05). The oxygenation indexes of patients in group E at admission and shock stage and on PID 3 to 7 [(371±16), (263±16), and (228±18) mmHg (1 mmHg=0.133 kPa)] were lower than (420±13), (327±13), and (281±17) mmHg of patients in group YM, respectively (t=5.650, 9.782, 4.856, P<0.05 or P<0.01). There were no statistically significant differences in levels of blood platelet count at admission and shock stage between patients in the 2 groups (t=0.038, 0.588, P>0.05), while the level of blood platelet count of patients in group E on PID 3 to 7 was significantly lower than that in group YM (t=6.636, P<0.05). (3) The days of seriously or critically ill and death rate of patients in group E were respectively longer or higher than those in group YM (Z=-2.303, χ2=13.676, P<0.05 or P<0.01).@*Conclusions@#In the case of the same tissue perfusion at shock stage, injuries in heart, liver, kidney, lung, and coagulation system in elderly patients with severe burns are more obvious than those in young and middle-aged patients, with more severe illness and higher mortality.
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Objective To investigate the molecular mechanism of retinoic acid in targeted treatment of craniopharyngioma by detecting the expression of RARαand PPARβ/δin craniopharyngioma cells and analyzing the correlation between the expression and effect of retinoic acid. Methods The expression of RARα and PPARβ/δ in craniopharyngioma cells from 31 patients cultured in vitro was quantified by reverse transcription-PCR. The inhibition rates of RA on craniopharyngioma with different expression of RARα and PPARβ/δ were detected by using MTT assay, and the correlation between the expression of RARα and PPARβ/δand the effect of RA was analyzed. Results 1. The RT-PCR results showed that the expression levels of PPARβ/δand RARα mRNA were different. Craniopharyngioma cells from 31 patients in primary culture were divided into three groups according the expression levels of nuclear receptor: PPARβ/δ>RARα group, RARα>PPARβ/δ group and RARα>>PPARβ/δ group. 2.MTT results showed that the inhibition rate of RARα>>PPARβ/δgroup was significantly higher than the other groups under same drug, the differences had statistical significance ( <0.01) . Conclusions The expression of PPARβ/δ, RARα can be used to evaluate the effect of RA in treatment of craniopharyngioma. The craniopharyngioma with low-expression of PPARβ/δ is more sensitive to RA. Targeting higher RARα or targeting lower PPARβ/δ is beneficial to the treatment of craniopharyngiomas.
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Objective To explore the relationship between the expression of XRCC1 and glioma. Methods Total of 26 samples of glioma were divided into 4 groups:gradeⅠ,gradeⅡ,gradeⅢand gradeⅣ. The expression of XRCC1 in 26 Gliomas tissues were examined using SP immunohistochemical staining.Results The positive staining of XRCC1 protein was localized in nucleus of tumor cells in Glioma. There was no correlation among them. The difference of XRCC1 expression among gradeⅠ~Ⅳ was not significant ( >0.05) .Conclusion The difference of XRCC1 expression among gradeⅠ~Ⅳ was not significant. The expression of XRCC1 was closely correlated with the effect of radiotherapy.