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Objective:To evaluate the effect of gastrogin on AMP-activated protein kinase (AMPK)/transient receptor potential anchor protein 1 (TRPA1) signaling pathway in rats with neuropathic pain.Methods:Thirty-six SPF-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-230 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation+ normal saline group (SHAM group), neuropathic pain+ normal saline group (NP group), and neuropathic pain+ gastrogin group (GAS group). Neuropathic pain was induced by chronic constrictive injury to sciatic nerve under 2% isoflurane anaesthesia. The sciatic nerve was only exposed but not ligated in SHAM group. Gastrogin 100 mg/kg was intraperitoneally injected for 14 consecutive days after developing the model in GAS group, while the equal volume of normal saline was given instead in SHAM and NP groups. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before developing the model (T 0) and 1, 3, 5, 7, 10 and 14 days after developing the model (T 1-6). The rats were anesthetized and sacrificed following the measurement of pain thresholds at T 4 and T 6. The lumbar segment (L 4-6) of the spinal cord was removed for determination of TRPA1 mRNA expression (by quantitative real-time polymerase chain reaction), expression of TRPA1, AMPK and p-AMPK (by Western blot), expression of TRPA1 (by immunofluorescence staining) and expression of tumor necrosis-alpha(TNF-α), interleukin-1beta(IL-1β) and c-fos (by immunohistochemistry). Results:Compared with SHAM group, MWT and TWL were significantly decreased at T 1-6, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was up-regulated, the expression of p-AMPK was down-regulated ( P<0.05), and no significant change was found in AMPK expression in NP group ( P>0.05). Compared with NP group, MWT at T 3-6 and TWL at T 2-6 were significantly increased, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was down-regulated, and p-AMPK expression was up-regulated ( P<0.05), and no significant change was found in AMPK expression in GAS group ( P>0.05). Conclusions:The mechanism by which gastrogin reduces neuropathic pain may be related to modulating the expression of the AMPK/TRPA1 signaling pathway in rats.
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ObjectiveTo explore the mediating effect of personality traits between job stress and mental symptoms among firefighters. Methods A total of 974 firefighters in Shandong Province were selected as the research subjects using convenience sampling method. The Job Stress Scale, Brief Version of Chinese Big Five Personality Inventory and Symptom Checklist 90 were used to investigate their job stress, personality traits and mental symptoms. Results Job stress was found at a relatively high level in 39.7% of the firefighters, and 19.2% of the firefighters had high level of neurotic personality trait tendency. The positive detection rate of mental symptoms was 9.9% among firefighters. The score of job stress of the firefighters had a positive correlation with neuroticism and the score of mental symptoms (all P<0.01). The score of job stress and mental symptoms of the firefighters had a negative correlation with the personality traits score of extraversion, openness, agreeableness, and conscientiousness (all P<0.05). Neuroticism played a partially positive mediating effect between job stress and mental symptoms, and its mediating effect accounted for 39.6% of the total effect. Extraversion, agreeableness and conscientiousness played a partially negative mediating effect between work stress and mental symptoms, and the mediating effect accounted for 12.0%, 17.8% and 8.4% of the total effect respectively. There was no mediating effect of openness between job stress and mental symptoms. Conclusion Neurotic personality trait may enhance the negative effects of work stress on firefighters' mental health.
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Objective:To evaluate the role of Sigma-1 receptor (Sigma-1R) in pentazoxine-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury in SH-SY5Y cells and the relationship with endoplasmic reticulum stress (ERS).Methods:The well-growing SH-SY5Y cells were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), OGD/R group (group O), OGD/R+ pentazoxin group (group OP) and OGD/R+ pintazoxin+ BD1047 group (group OPB). The cells in group C were normally cultured. In O group, OP group and OPB group, the culture medium was replaced with EBSS medium, and then the cells were cultured in an incubator of 5% CO 2-95% N 2 at 37 ℃ for 4 h, then replaced with DMEM/F12 medium containing 10% fetal bovine serum for restoration of O 2-glucose supply for 18 h, and in addition pentazoxin (final concentration 10 μmmol/L) was added during restoration in OP group, and pentazoxin (final concentration 10 μmmol/L) and Sigma-1R blocker BD1047 (final concentration 20 μmol/L) were added during restoration in OPB group. The apoptosis rate was detected by flow cytometry at the end of restoration, and the expression of Sigma-1R, C/EBP homologous protein (CHOP), phosphorylated inositol-requiring enzyme 1 (p-IRE1), spliced X-box binding protein 1 (XBP1s), and activated caspase-3 (c-cas-3) was detected by Western blot. Results:Compared with group C, the apoptosis rate was significantly increased, and the expression of CHOP and p-IRE1 was up-regulated in O group, OP group and OPB group, the expression of XBP1s and c-cas-3 was significantly up-regulated in O group and OPB group ( P<0.05), and no significant change was found in the expression of Sigma-1R, XBP1s and c-cas-3 in OP group ( P>0.05). Compared with O group, the apoptosis rate was significantly decreased, the expression of Sigma-1R was up-regulated, and the expression of CHOP, p-IRE1, XBP1s and c-cas-3 was down-regulated in OP group ( P<0.05). Compared with OP group, the apoptosis rate was significantly increased, the expression of Sigma-1R was down-regulated, and the expression of CHOP, p-IRE1, XBP1s and c-cas-3 was up-regulated in OPB group ( P<0.05). Conclusions:Sigma-1R is involved in pentazoxine-induced reduction of OGD/R injury in SH-SY5Y cells, and the mechanism may be related to inhibition of endoplasmic reticulum stress.
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Objective: To study the relationship between compassion fatigue and overall happiness among firefighters, and the mediating role of psychological stress and positive psychological capital. Methods: A total of 894 firefighters in Shandong Province were selected as the research subjects using purposive sampling method. Questionnaires including the Professional Quality of Life Scale, the Military Psychological Stress Self-Assessment Scale, the Positive Psychological Capital Scale, and the Overall Happiness Scale were used for data collection. Results: The scores of compassion fatigue, psychological stress, positive psychological capital and overall happiness among firefighters were (40.1±13.5), (13.9±3.9), (133.0±26.4) and (84.9±15.2), respectively. There were correlations between compassion fatigue, psychological stress, positive psychological capital and overall happiness (all P<0.01). Psychological stress partially mediated the relationship between compassion fatigue and overall happiness, and the mediating effect accounted for 27.0% of the total effect. Positive psychological capital moderated the front half path and the direct path between compassion fatigue and overall happiness (all P<0.01). Conclusion: Compassion fatigue can directly or indirectly affect the firefighters' overall happiness. Psychological stress plays a partial mediating role and positive psychological capital plays a moderating role between compassion fatigue and overall happiness.
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Deepening the teaching reform of medical education is the strategic initiatives to improve the quality of medical talents in the new period. At present, curriculum teaching reform and the certification work in medical colleges and universities is still in the initial exploration stage. Although some achievements have been made, there are still some prominent problems, such as insufficient teacher drive, inaccurate orientation for the curriculum, improper ways of teaching, lack of organizational and appraising system. Xuzhou Medical University takes the declaration and accreditation of educational reform courses as the forerunner, and takes many measures to stimulate teachers' endogenous motivation of educational reform, innovate the organizational system of educational reform courses evaluation, establish progressive evaluation and accreditation management, formulate evaluation standards of educational reform courses, and form a closed loop of multiple evaluation and feedback. It not only effectively improves the quality of teaching and further promote the curriculum reform process, but also provides reference for the development and promotion of curriculum teaching reform in other colleges and universities.
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Objective To analyze the epidemiological characteristics and risk factors of bronchial asthma (BA) complicated with recurrent respiratory tract infection (RRTI) in children in Hebei District, Tianjin City, and to provide a reference for the prevention and treatment of children with BA complicated with RRTI. Methods The stratified cluster sampling method was adopted to randomly select 428 children with BA hospitalized in Hebei District of Tianjin as the study subjects. The routine deep sputum culture and etiological examination were carried out. The children with RRTI were included in the experimental group (n=84), and the children without RRTI (n=344) were included in the control group. A self-designed questionnaire was used to investigate gender, age, smoking proportion of family members, use of antibiotics 3 times or more a year, and family history of allergy. The risk factors of BA combined with RRTI were analyzed by logistic regression. Results RRTI occurred in 84 of 428 children with BA, and the incidence rate was 19.63% (84/428). The proportion of BA complicated with RRTI in children aged 6 months to 2 years was higher than that in other age groups (χ2=6.213, P0.05). The proportion of family smoking, the use of antibiotics 3 times or more a year, and the proportion of family allergy history in the experimental group were significantly higher than those in the control group (P<0.05). Logistic regression analysis showed that family smoking, antibiotic use 3 times or more a year and family history of allergy were independent risk factors for BA complicated with RRTI (P<0.05). Conclusion The incidence of BA complicated with RRTI in children in Hebei District, Tianjin City is high, and the age of high incidence is 6 months to 2 years old. The proportion of family smoking, the use of antibiotics three times or more a year, and the proportion of family allergy history are the high-risk factors for the occurrence of BA complicated with RRTI in children in Hebei District, Tianjin City.
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ObjectiveTo investigate the association of gene mutations in the pre-C, C, and basic core promoter (BCP) regions of hepatitis B virus (HBV) with traditional Chinese medicine (TCM) syndrome types in patients with chronic hepatitis B (CHB). MethodsA retrospective analysis was performed for the clinical data of CHB patients who were diagnosed and treated at the outpatient service and ward of Spleen, Stomach, and Hepatobiliary Department, The First Affiliated Hospital of Henan University of Chinese Medicine, from November 2014 to June 2018. Related clinical data were collected and recorded, including general information, HBV serological markers, HBV gene mutations, and information obtained by four TCM diagnostic methods. Syndrome differentiation and typing were performed for each patient with reference to the criteria for TCM syndrome differentiation of viral hepatitis, and the association of gene mutation in the pre-C, C, and BCP regions of HBV with TCM syndrome types was analyzed. The chi-square test was used for comparison of categorical data between groups, and the Kruskal-Wallis H test was used for comparison of continuous data between multiple or two groups. ResultsA total of 235 patients with CHB were enrolled, among whom 101 had internal retention of damp-heat, 88 had stagnation of liver Qi and spleen deficiency, 17 had blood stasis obstructing the collaterals, 19 had liver-kidney Yin deficiency, and 10 had spleen-kidney Yang deficiency. There were significant differences in sex, age, and course of disease between the patients with different TCM syndrome types (χ2=17.389, H=173.280, H=86.520, all P＜0.01), and there was a significant difference in age between the CHB patients with gene mutations in the pre-C, C and BCP regions of HBV (H=30.150, P＜0.001). There was a significant difference in the distribution of TCM syndrome types between the CHB patients with gene mutations in the pre-C, C and BCP regions of HBV (χ2=58.117, P＜0.001), and internal retention of damp-heat and stagnation of liver Qi and spleen deficiency were major TCM syndrome types accounting for 80.43%. The patients with internal retention of damp-heat tended to have A1762T and G1764A mutations, and those with stagnation of liver Qi and spleen deficiency tended to have G1896A, A1762T, and G1764A mutations; G1764A mutation was often observed in the patients with blood stasis obstructing the collaterals or liver-kidney Yin deficiency, and I97L mutation was often observed in the patients with spleen-kidney Yang deficiency. ConclusionGene mutations in the pre-C, C, and BCP regions of HBV are associated with TCM syndrome types in CHB patients, and internal retention of damp-heat and stagnation of liver Qi and spleen deficiency are the most common TCM syndrome types. I97L mutation is often observed in patients with spleen-kidney Yang deficiency.
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Objective:To investigate the role of Sestrin2 overexpression in regulating mitochondrial fission and its mechanism in human neuroblastoma SH-SY5Y cell model of glucose and oxygen deprivation/recovery (OGD/R). Methods:(1) SH-SY5Y cells were divided into normal control group, OGD/R group, Vector group, and Sestrin2 overexpression group; Sestrin2 overexpression or empty vector stable cell lines in the Sestrin2 overexpression group and Vector group were constructed by lentivirus infection; cells in the later 3 groups were subjected to oxygen-glucose deprivation (OGD) for 4 h followed by restoration of O 2 supply for 18 h. The cell survival rate was detected by cell counting kit (CCK)-8 assay. The protein levels of Sestrin2, dynamin-related protein 1 (Drp1), mitochondrial fission protein 1 (Fis1), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), Kelch-like ECH-related protein 1 (Keap1) in the cytoplasm and nuclear factor E2-related factor (Nrf2) in the nucleus were detected by Western blotting. The mitochondria ultrastructure was observed by transmission electron microscope. The Nrf2 nuclear translocation was detected by immunofluorescence staining. (2) Cell lines with Sestrin2 overexpression were divided into Sestrin2 overexpression group, Brusatol+ Sestrin2 overexpression group, and DMSO+ Sestrin2 overexpression group. Cells in the Brusatol+ Sestrin2 overexpression group were pretreated with normal medium containing Brusatol (Keap1/Nrf2 pathway inhibitor, final concentration: 100 nmol/L) for 4 h before OGD/R; cells in the DMSO+ Sestrin2 group were pretreated with normal medium containing DMSO (final volume fraction: 0.1%) for 4 h before OGD/R. Cells in these groups were then subjected to OGD for 4 h followed by restoration of O 2 supply for 18 h. The protein levels of Drp1, Fis1, Keap1 in the cytoplasm, and Nrf2 in the nucleus were measured by Western blotting. Results:(1) As compared with those in the OGD/R group, cells in the Sestrin2 overexpression group had significantly increased survival rate (61.33%±1.15% vs. 81.00%±3.00%), significantly up-regulated Bcl-2/Bax ratio (0.467±0.006 vs. 0.880±0.010), significantly decreased Drp1, Fis1 and cytoplasmic Keap1 protein levels (1.089±0.033 vs. 0.865±0.014; 0.829±0.009 vs. 0.350±0.007; 0.967±0.017 vs. 0.881±0.024), and significantly up-regulated nuclear Nrf2 protein level (0.627±0.025 vs. 0.957±0.015, P<0.05). The mitochondrial structure in the Sestrin2 overexpression group under electron microscope was more complete than that in the OGD/R group, and obvious nuclear translocation of Nrf2 was noted. (2) As compared with the Sestrin2 overexpression group, Brusatol+ Sestrin2 overexpression group had significantly decreased nuclear Nrf2 protein level (0.920±0.013 vs. 0.627±0.035), and statistically increased Drp1 and Fis1 protein levels (0.994±0.020 vs. 1.084±0.005; 0.728±0.010 vs. 0.906±0.022, P<0.05). Conclusion:Sestrin2 overexpression could suppress mitochondrial fission, reduce cell apoptosis, and attenuate OGD/R injury of SH-SY5Y cells by activating Keap1/Nrf2 pathway via down-regulating cytoplasmic Keap1 protein level and promoting Nrf2 nuclear translocation.
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Objective To investigate the protective role of Sestrin2 overexpression in hypoxia and re-oxygenation injury (H/R) injury of hippocampal neurons and its mechanism in rats.Methods Neurons were enzymatically isolated from hippocampi of newborn Sprague-Dawley rats (less than 24 h old) and culturedinvitro. These neurons were randomly assigned into 4 groups (n=20) using a random number table: control group, H/R group, vector group and Sestrin2 overexpression group. The hippocampal neurons were seeded in 6-well plates at a density of 2×104 cells/mL; neurons in the latter two groups were transfected with lentiviruses containing empty vector andSestrin2overexpressed genes, respectively; the hippocampal neurons in the later three groups were subjected to oxygen-glucose deprivation (OGD) for 6 h followed by restoration of O2 supply for 20 h. The reactive oxygen species (ROS) content was detected by Reactive Species Assay Kit, and the ATP concentration was detected by ATP Assay Kit. Cell apoptosis rates were measured by flow cytometry. The levels of Sestrin2, dynamin-related protein 1 (Drp1), Fis1, and apoptosis-related proteins Bcl-2, Bax and cytochrome C (Cyt C) were measured by Western blotting. The ratio of Bcl-2 to Bax was calculated. The ultrastructure of mitochondria was observed by transmission electron microscopy.Results As compared with control group, H/R group had significantly lower ATP concentration, Bcl-2 protein expression and ratio of Bcl-2 to Bax ([11.15±0.42] nmol/mg proteinvs. [5.30±0.39] nmol/mg protein; 2.20±0.26vs. 0.91±0.02; 6.46± 0.41vs. 1.04±0.05), statistically higher average fluorescence intensity of ROS and cell apoptosis rate (152.41±17.38vs. 1530.00±14.69; 3.77％±0.74％vs. 56.57％±1.35％), and significantly higher protein levels of Sestrin2, Drp1, Fis1, Bax and Cyt C (0.66±0.06vs. 1.11±0.20; 0.48±0.03vs. 1.16±0.07; 1.14± 0.09vs. 2.47±0.09; 0.34±0.03vs. 0.88±0.04; 0.17±0.03vs. 0.30±0.03,P<0.05); what's more, the structure of mitochondria was obviously destroyed in I/R group. As compared with H/R group, Sestrin2 overexpression group had significantly increased ATP concentration, Sestrin2 and Bcl-2 protein expressions and ratio of Bcl-2 to Bax ([8.95±0.27] nmol/mg protein; 2.67±0.07; 1.80±0.19; 3.95±0.28), significantly lower average fluorescence intensity of ROS and cell apoptosis rate (337.27±15.32; 10.33％±2.60％), and statistically lower protein levels of Drp1, Fis1, Bax and Cyt C (0.43±0.02; 1.11±0.08; 0.45± 0.02; 0.17±0.02,P<0.05); the structure of mitochondria was relatively completed in Sestrin2 overexpression group.Conclusion Sestrin2 overexpression can inhibit mitochondrial fission, reduce accumulation of reactive oxygen species, improve mitochondrial energy metabolism and block mitochondria mediated apoptosis pathway, thereby alleviating I/R injury of rat hippocampal neurons.
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Objective To evaluate the role of mitochondrial calcium uniporter ( MCU) in mitoph-agy in SH-SY5Y cells subjected to oxygen-glucose deprivation and restoration (OGD∕R). Methods SH-SY5Y cells were cultured in vitro, seeded in 96-well plates at a density of 2×105cells∕ml, and randomly di-vided into 4 groups (n=6 each) using a random number table method: control group (group C), group OGD∕R, OGD∕R plus MCU inhibitor group ( group OGD∕R + Ru360) and MCU inhibitor group ( group Ru360) . Cells were cultured in normal culture medium in group C. Cells were subjected to O2-glucose dep-rivation for 6 h followed by restoration of O2-glucose supply for 24 h in group OGD∕R. In group OGD∕R+Ru360, Ru360 at a final concentration of 10 μmol∕L was added at 30 min before O2-glucose deprivation, and the other treatments were similar to those previously described in group OGD∕R. Ru360 was added at a final concentration of 10 μmol∕L, and 30 min later cells were cultured under normoxic conditions in group Ru360. At 24 h of restoration of O2-glucose supply, cell counting kit-8 assay was used to detect the cell survival rate, JC-1 assay was used to detect mitochondrial membrane potential ( MMP ) , the ultrastructure of cells was observed with a transmission electron microscope, and the expression of p62, Tom20 and Bec-lin-1 was detected by Western blot. Results Compared with group C, no significant change was found in each parameter in group Ru360 ( P>0. 05) , the cell survival rate and MMP were significantly decreased, the expression of Tom20 and p62 was down-regulated, Beclin-1 expression was up-regulated (P<0. 01), the mitochondria swelled, and mitochondrial autophagosomes were increased in group OGD∕R. Compared with group OGD∕R, the cell survival rate and MMP were significantly increased, the expression of Tom20 and p62 was up-regulated, Beclin-1 expression was down-regulated (P<0. 01), the mitochondrial mor-phology kept well, and mitochondrial autophagosomes were decreased. Conclusion MCU is involved in the process of mitophagy in SH-SY5Y cells subjected to OGD∕R.
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Objective To evaluate effect of dexmedetomidine on mitochondrion-dependent apoptosis during hypoxia-reoxygenation (H/R) injury to hippocampal neurons of rats.Methods The primarily cultured hippocampal neurons of Sprague-Dawley rats were divided into 4 groups (n =40 each) using a random number table method:control group (C group),vehicle group (V group),H/R group and dexmedetomidine group (D group).Hippocampal neurons were subjected to oxygen-glucose deprivation followed by restoration of oxygen supply to establish the model of H/R injury.Dexmedetomidine 1 μmol/L was added at 6 h of reoxygenation in D group.The viability of neurons was measured by methyl thiazolyl tetrazolium assay at 20 h of reoxygenation.The ultrastructure of mitochondria was observed by transmission electron microscopy.The expression of cytochrome c (Cyt c),caspase-3,Fis1 and Drp1 was detected by Western blot.The neuronal apoptosis was detected by flow cytometry,and apoptosis rate was calculated.Results Compared with C group,no significant change was found in the viability of neurons in group V (P＞0.05),and the viability of neurons was significantly decreased,the apoptosis rate was increased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was up-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was accentuated in H/R and D groups.Compared with H/R group,the viability of neurons was significantly increased,the apoptosis rate was decreased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was down-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was significantly attenuated in D group.Conclusion The nechanism by which dexmedetomidine reduces the H/R injury to hippocampal neurons is related to inhibiting mitochondrion-dependent apoptosis in rats.
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Objective To investigate the efficacy of high frequency jet ventilation via the nasopha?ryngeal catheter in assissting ventilation in patients with obstructive sleep apnea hypopnea syndrome ( OS?AHS) undergoing gastroscopy. Methods Eighty patients with OSAHS of both sexes, aged 40-64 yr, weighing 65-99 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with an apnea?hy?popnea index 20-40 events∕h, scheduled for elective painless gastroscopy, were divided into control group and test group using a random number table, with 40 patients in each group. In control group, a nasopha?ryngeal catheter 6.0-7.0 mm in internal diameter was inserted, and oxygen was inhaled at 4 L∕min through the catheter. In test group, a nasopharyngeal catheter 4.0 mm in internal diameter was inserted, and a high frequency jet ventilator was connected ( inspiratory∕expiratory ratio 1. 0 ∶ 1. 5, frequency 150 bpm, peak pressure 0.4 kPa, tidal volume 180 ml) . Anesthesia was maintained with propofol in both groups. The oc?currence of hypoxemia during ventilation, and peak value of partial pressure of end?tidal CO2 before induc?tion of anesthesia and during ventilation, and occurrence of chin lift, mask ventilation, and epistaxis after insertion of the catheter during operation were recorded. Results Compared with control group, the inci?dence of hypoxemia, peak value of partial pressure of end?tidal CO2 during ventilation, and incidence of chin lift, mask ventilation and epistaxis during operation were significantly decreased in test group ( P<0.05) . Conclusion High frequency jet ventilation via the nasopharyngeal catheter can be safely and effec?tively used to assisst ventilation in patients with OSAHS undergoing gastroscopy.
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Objective To evaluate the effect of propofol on mitochondrial fission in a rat hippocampal neuron model of hypoxia/reoxygenation (H/R) injury.Methods Primarily cultured hippocampal neurons of neonatal Sprague-Dawley rats were randomly divided into 4 groups (n =42 each) using a random number table:control group (group C);vehicle group (group V);H/R group;H/R+propofol group (group H/R+P).In group V,H/R was not produced,the vehicle dimethyl sulfoxide with the final concentration of 0.01％ was added,and the cells were then incubated for 6 h.In group H/R,the hippocampal neurons were subjected to oxygen-glucose deprivation for 6 h followed by 20 h reoxygenation.In group H/R+P,propofol with the final concentration of 1 μmol/L was added at 6 h of hypoxia.At 20 h of reoxygenation,the cell apoptosis (using flow cytometry),Ca2+ concentrations in cytoplasm (with the laser scanning confocal microscope),calcineurin (CaN) activities (by enzyme-linked immunosorbent assay),and expression of mitochondrial fission proteins dynamin-related protein 1 (Drp1) and fission 1 (Fis1),and apoptosis-related proteins cytochrome c (Cyt c) and apoptosis-inducing factor (AIF) (by Western blot) were measured.The apoptotic rate was calculated.Results Compared with group C,the apoptotic rate,Ca2+concentrations,CaN activities,and expression of Drp1,Fis1,Cyt c and AIF were significantly increased in H/R and H/R+P groups (P＜0.05),and no significant changes were found in the parameters mentioned a2+.bove in group V (P＞0.05).Compared with group H/R,the apoptotic rate,Ca+ concentrations,CaN activities,and expression of Drp1,Fis1,Cyt c and AIF were significantly decreased in group H/R+P (P＜0.05).Conclusion Propofol can reduce the H/R injury to rat hippocampal neurons through inhibiting mitochondrial fission.
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Objective To explore the relationship between life events,efficacy of self learning and learning burnout of Left-behind middle school Students.Methods 526 left-behind middle school students and 860 non -left-behind students were tested by means of scales of life events,academic self-efficacy and learning burnout.Results The degree of interpersonal relationship,learning pressure,health adaptation,life events,self-efficacy of learning ability,learning behaviors,and learning,sense of alienation between teacher and students,physiological depletion,and learning burnout of left-behind middle school students were lower than non left-behind students,and were statistically significant.The interpersonal relationship(r=0.270,P＜0.001),learning pressure(r=0.289,P＜ 0.001),being punished(r=0.242,P＜0.001),health adaptation(r=0.301,P＜0.001) and others (r=0.322,P＜ 0.001) were positively related to learning burnout.Self-efficacy of learning ability(r=-0.334,P＜0.01) and learning behaviors(r=-0.157,P＜0.001) were significantly and negatively related.Interpersonal relationship (r=-0.092,P＜0.05),learning pressure (r=-0.123,P＜0.01),being punished (r=-0.178,P＜0.001) and others (r=-0.254,P＜ 0.001) were negatively related to self-efficacy of learning ability.Being punished (r=-0.109,P＜ 0.05)and others(r=-0.209,P＜0.001) were significantly and negatively related to self-efficacy of learning behaviors.Self-efficacy,health adaptation,learning pressure,self-efficacy of learning behavior and others all entered the regression equation.Path analysis showed that there were five paths that have significant influences on learning burnout.Conclusion The levels of response to emergency of life events and enhance the academic self-efficacy of leftbehind students can be improved to reduce and prevent learning burnout.
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Objective To evaluate the role of mitochondrial fission in hypoxia-reoxygenation (H/ R) injury to hippocampal neurons of rats.Methods Primarily cultured hippocampal neurons obtained from newborn Wistar rats were randomly divided into 5 groups (n =60 each) using a random number table: normal group (N group), vehicle group (V group), H/R group, H/R + vehicle group (H/R + V group),and mitochondrial division inhibitor group (group M).The cells were cultured in normal culture medium in group N.Dimethyl sulfoxide (DMSO) was added to the culture medium with the final concentration ＜ 0.1％, and the cells were incubated for 40 min in group V.The cells were subjected to 6 h hypoxia, followed by 20 h reoxygenation in H/R, H/R+V and M groups.DMSO was added to the culture medium with the final concentration ＜0.1％ at 40 min before hypoxia in group H/R+V.In group M, mitochondrial division inhibitor Mdivi-1 50 mmol/L (dissolved in DMSO, DMSO concentration ＜0.1％) was added to the culture medium at 40 min prior to hypoxia.Mito Tracker staining was used to examine mitochondrial morphology.Western blot was used to measure the expression of mitochondrial dynamin-related protein 1 (Drp1), mitofusin 2 (Mfn2) , peroxisome proliferator activated receptor γ coactivator 1α (PGC-1α), nuclear respiratory factor-1 (NRF-1), and mitochondrial transcription factor A (TFAM).Multifunctional microplate reader and fluorescent microscope were used to detect the mitochondrial reactive oxygen species (ROS) level.The flow cytometer was used to detect the apoptosis in hippocampal neurons.Apoptosis rate was calculated.Results Compared with group N, the expression of Drp1, PGC-1α, NRF-1 and TFAM was significantly up-regulated, the ROS content and apoptosis rate were increased, and the expression of Mfn2 was down-regulated in group H/R (P＜0.05).Compared with group H/R, the expression of Drp1 was significantly down-regulated, the ROS content and apoptosis rate were decreased, and the expression of Mfn2, PGC-1α, NRF-1 and TFAM was up-regulated in group M (P＜0.05).Conclusion Mitochondrial fission is involved in H/R injury to hippocampal neurons of rats.
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Objective To evaluate the role of mitochondrial fission in anoxia-reoxygenation injury to rat hippocampal neurons.Methods Neurons were enzymatically isolated from hippocampi of newborn Sprague-Dawley rats (less than 24 h old).The primary hippocampal neurons were cultured and seeded in 25 mm × 25 mm culture flasks at a density of 7 × 105/ml.The cultured neurons were randomly assigned into 3 groups (n =18 each) using a random number table:control group (C group),anoxia-reoxygenation group (I/R group),and mitochondrial fission inhibitor mdivi-1 group (M group).In group I/R,the vehicle dimethyl sulfoxide (DMSO,final concentration ＜ 0.1％) was added prior to anoxia and the cells were then incubated for 40 min.In group M,mdivi-1 (dissolved in DMSO,final concentration of DMSO ＜ 0.1％) was added prior to anoxia and the cells were then incubated for 40 min.The hippocampal neurons were subjected to oxygen-glucose deprivation (OGD) for 6 h followed by restoration of O2 supply for 20 h.After 20 h of reoxygenation,the level of reactive oxygen species (ROS) (by ELISA),cell apoptosis (using flow cytometry),and expression of mitochondrial fission protein Drp1,Bcl-2 and Bax (by Western blot) were measured.The apoptosis rate and the ratio of Bcl-2 to Bax were calculated.Results Compared with C group,ROS content and apoptosis rate were significantly increased,the expression of Drp1 and Bax was up-regulated,the expression of Bcl-2 was down-regulated,and the ratio of Bcl-2 to Bax was decreased in I/R group (P ＜ 0.05).Compared with I/R group,ROS content and apoptosis rate were significantly decreased,the expression of Drp1 and Bax was down-regulated,the expression of Bcl-2 was up-regulated,and the ratio of Bcl-2 to Bax was increased in M group (P ＜ 0.05).Conclusion Mitochondrial fission is involved in anoxia-reoxygenation injury to rat hippocampal neurons via mitochondria-mediated apoptotic pathway.
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Objective To analyze the pattern of normal bone marrow on whole body diffusionweighted imaging (WB-DWI) and its influence factors.MethodsA total of 98 healthy volunteers (male 47 and female 51). All volunteers underwent WB-DWI at 1.5 T MR scan. The ADC value,the signal intensity on DWI obtained with a b value of 800 s/mm2 ( SIDWI ),and the signal intensity on short time inversion recovery images ( SISTIR ) of thoracic vertebrae,lumber vertebrae,bilateral head of femur,bilateral neck of femur,bilateral superior segment of femur,bilateral inferior segment of femur,bilateral ilium,bilateral head of humerus,bilateral scapula were measured and compared with ANOVA test and StudentNewmar-Keuls test.The normal appearance of bone marrow on WB-DW[ was assessed. The relationship between the SIDwI and the ADC,and the SIDWI and the SISTIR of bone marrow were analyzed.The ADC of bone marrow between male and femaIe were compared. Spearman correlation analysis was performed for different age groups.Results( 1 ) Bone marrow signal intensity was different among 98 healthy volunteers.Bone marrow in 69 healthy volunteers (female 24,male 45 ) showed low to intermediate signal intensity,whereas in the remaining 29 healthy volunteers (female 27,male 2) showed high signal intensity.(2) The SIDWI of thoracic vertebrae ( median 44.54),lumber vertebrae ( median 35.01 ),head of femur ( median 13.61 ),neck of femur ( median 16.00),superior segment of femur ( median 21.45 ),ilium ( median 25.77),head of humerus (median 18.35),scapula (median 36.12) was positively correlated with the ADC [ (0.55 ±0.08) × l0-3,(0.53 ±0.08) × 10-3,(0.30 ± 0.10) × 10-3,(0.42 ± 0.16) × 10-3,(0.74±0.14) ×l0- 3,(0.49±0.10) ×10-3,(0.36±0.13) ×10-3,(0.49±0.11) × 10-3mm2/s]and the SISTIR ( median 61.81,64.99,53.27,69.08,73.10,66.35,73.16,79.81 ),r =0.513 and 0.695,0.741 and 0.764,0.443 and 0.489,0.641 and 0.656,0.510 and 0.648,0.475 and 0.715,0.366 and 0.446,0.437 and 0.739 ;P ＜ 0.01. (3) There was significant difference of the ADC of bone marrow in different bone,F =138.69,P ＜ 0.01. Student-Newman-Keuls test revealed that no significant difference was found in the ADC between thoracic vertebrae and lumbar vertebrae,ilium and scapula,head of humerus and inferior segment of femur ( P ＞ 0.05 ),and significant difference was found in the ADC values between the remaining two groups ( P ＜ 0.05 ). The bones associated with decreasing ADC values were superior segment of femur,thoracic vertebrae and lumber vertebrae,ilium and scapula,neck of femur,head of humerus,head of femur and inferior segment of femur. ( 4 ) The ADC values of bone marrow of female subjects in thoracic vertebrae [ (0.59 ±0.07) × 10-3 mm2/s],lumber vertebrae [ (0.58 ±0.06) × 10 -3 mm2/s],head of femur ( median 0.33 × 10 -3 mm2/s),neck of femur ( median 0.53 × 10 -3 mm2/s),superior segment of femur ( median 0.81 × 10-3 mm2/s),inferior segment of femur ( median 0.32 ×10-3 mm2/s),ilium [ (0.52 ± 0.09 ) × 10-3 mm2/s ],head of humerus (median 0.42 × 10-3 mm2/s),scapula [ (0.53 ± 0.09) × 10-3 mm2/s] were significantly higher than those of male subjects [ (0.51 ±0.07) × 10-3,(0.48 ±0.07) × 10-3,median 0.23 × 10-3,median 0.31 × 10-3,median 0.66 × 10-3,median 0.23 × 10-3,(0.46 ±0.10) × 10-3,median 0.27 × 10-3,(0.45 ±0.11 ) × 10 3mm2/s].(5)There was significant negative correlation between the ADC values of bone marrow and age in thoracic vertebrae,lumber vertebrae,head of femur,neck of femur,superior segment of femur,ilium,head of humerus for female subjects,r =-0.549, -0.629, -0.329, -0.524, -0.338, -0.548 and -0.416,respectively,P ＜ 0.05.There was no significant correlation between ADC values and age in inferior segment of femur and scapula for female subjects and all the regions for male subjects ( P ＞ 0.05 ).Conclusions The ADC and the SIsTIR of bone marrow correlates with the SIDW1.The ADC values of bone marrow is affected by age and sex,and is different for different bones.
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ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.
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BACKGROUND: Both abnormal permeability of ionic channel and disturbance of ionic balance between inside and outside nerve cell are key factors for ischemic brain injury after ischemia. Depolarization induced by activation of sodium channel is starting link for cerebral ischemic injury.OBJECTIVE: To study the effects of droperidol on persistent sodium channel currents of pyramidal cell in hippocampal CA1 area of rats with cerebral ischemia with patch clamp technique so as to analyze whether droperidol can protect cerebral ischemic injury.DESIGN: Randomized controlled animal study.SETTING: Department of Anesthesiology of the Sixth People's Hospital Affiliated to Shanghai Jiaotong University and Department of Anesthesiology of the First People's Hospital Affiliated to Shanghai Jiaotong University.MATERIALS: The experiment was carried out at the Department of Anesthesiology of the First People's Hospital Affiliated to Shanghai Jiaotong University from April 2002 to April 2003. Totally 14 SD rats, aging 10-14days, without ablactation, were selected. Two cells in hippocampal CA1area of each rat were collected, totally 28 cells were divided into 4 groups:ischemic control group, 3 μmol/L droperidol group, 10 μmol/L droperidol group and 30 μmol/L droperidol group, with 7 cells in each group.METHODS: Pyramidal cells in hippocampal CA1 area were separated with digested enzyme method, and ischemic model of neuron was established through hypoxia and no sugar method. Cells were selected with the following conclusion criteria: well adherent wall, triangle or starry shape,bright soma, well refraction, obvious apophysis, steady plasma, and transparent nucleolus. Y-tube system was used for rapid medication. 3, 10 and 30 μmol/L droperidol were given to rats in 3, 10 and 30 μmol/L droperidols respectively, but rats in ischemic control group were not given any medicine. Whole-cell patch-clamp was used to recorded basic value of persistent sodium currents and changes of sodium channel currents during 3-minute and 5-minute ischemia.MAIN OUTCOME MEASURES: ① Record of normal persistent sodium current of neuron in cerebral hippocampal CA1 area; ② Record of persistent sodium current of neuron in cerebral hippocampal CA1 area during cerebral ischemia; ③ Effect of droperidol in various concentrations on persistent sodium current of neuron in cerebral hippocampal CA1 area during cerebral ischemia.RESULTS: Totally 28 cells in cerebral hippocampal CA1 area of 14 rats were entered the final analysis. ① Record of normal persistent sodium current of neuron in cerebral hippocampal CA1 area: 0.5 mmol/L CdCl2 calcium channel blocking agent and 20 mmol/L TEA kalium channel blocking agent were used to perform 400 ms square-wave stimulation under -105 mV claw voltage and -30 mV stimulated voltage. Introversion current,slight, late activation and lasting for a long time, was recorded and deter mined as persistent sodium currents by blocking toxin of puffer fish. ② Record of persistent sodium current of neuron in cerebral hippocampal CA1 area during cerebral ischemia: After 3-minute ischemia, persistent sodium currents in ischemic control group was increased as (1.60±0.21) times as that in normal group, and was (2.87 ±0.45) times after 5-minute ischemia. The difference was significant (P ＜ 0.05). ③ Effect of droperidol at various concentrations on persistent sodium current of neuron in cerebral hippocampal CA1 area during cerebral ischemia: Basic values of persistent sodium currents were (77.42±15.17) pA, (87.44±21.56) pA, (84.13±20.06) pA and (80.22±19.30) pA in ischemic control, 3, 10 and 30 μmol/L droperidol groups respectively, and the differences among groups were not significant. After 5-minute ischemia, values of persistent sodium currents were (105.36±17.16) pA, (94.74±18.88) pA and (84.88±13.94) pA in 3, 10 and 30 μmol/L droperidol groups respectively, which were obviously lower than that in the ischemic control group (218.31±29.34) pA.CONCLUSION: Persistent sodium currents increase under -105 mV claw voltage and -30 mV stimulated voltage during cerebral ischemic injury. Droperi dol can protect neuron by inhibiting the increase of persistent sodium current.
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AIM: To study the effects of droperidol on the enhancement of persistent sodium currents induced by in vitro ischemia-like condition in isolated rat CA1 paramidal neurons. METHODS: Whole-cell patch-clamp recordings were made from enzymatically isolated rat CA1 hippocampal paramidal neurons. Ischemia was induced by oxygen and glucose deprivation. RESULTS: All of 3, 10 ,and 30 ?mol?L -1 of droperidol significantly inhibited the enhancement of persistent sodium currents induced by ischemia, but the different concentrations did not show significant difference. CONCLUSION: Droperidol in clinical concentration can inhibit the enhancement of persistent sodium current induced by ischemia.