Role of GRSF1 in cerebral ischemia-reperfusion injury in mice: relationship with ferroptosis / 中华麻醉学杂志

Objective:To evaluate the role of G-rich RNA sequence binding factor 1 (GRSF1) in cerebral ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Twenty-four clean-grade male C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), cerebral I/R group (IR group), cerebral I/R+ GRSF1 overexpression group (IR+ LV-GRSF1 group), and cerebral I/R+ GRSF1 overexpression+ glutathione peroxidase 4 (GPX4) inhibitor group (IR+ LV-GRSF1+ RSL3 group). The model of middle cerebral artery occlusion was developed by thread-occlusion method in anesthetized animals. In IR+ LV-GRSF1 group, GRSF1-overexpressed lentivirus 2 μl was injected into the lateral ventricle at 7 days before the development of the model. GPX4 inhibitor RSL3 5 mg/kg was intraperitoneally injected for 2 consecutive days before the development of the model in IR+ LV-GRSF1+ RSL3 group. After 24 h of reperfusion, the percentage of cerebral infarction volume was determined by TTC assay, the survival neurons in ischemic area were detected by Nissl staining, and brain tissues in ischemic area were obtained for determination of the expression of p16, p21(markers of senescence) and tumor necrosis factor-alpha (TNF-α, senescence-associated secretory phenotype) mRNA (by quantitative real-time polymerase chain reaction), contents of malondialdehyde (MDA), superoxide dismutase(SOD) and glutathione (GSH) (by enzyme-linked immunosorbent assay) and expression of GRSF1, GPX4, Acyl-CoA synthetase long-chain family member 4 (ACSL4) and ferritin (by Western blot). Results:Compared with Sham group, the percentage of cerebral infarction volume was significantly increased, the count of viable neurons was decreased, the expression of p16, p21 and TNF-α mRNA in ischemic brain tissues was up-regulated, SOD and GSH contents were decreased, the MDA content was increased, the expression of GRSF1 and GPX4 was down-regulated, and the expression of ACSL4 and ferritin was up-regulated in IR group ( P<0.05). Compared with IR group, the percentage of cerebral infarction volume was significantly decreased, the count of viable neurons was increased, the expression of p16, p21 and TNF-α mRNA in ischemic brain tissues was down-regulated, SOD and GSH contents were increased, the MDA content was decreased, the expression of GRSF1 and GPX4 was up-regulated, and the expression of ACSL4 and ferritin was down-regulated in IR+ LV-GRSF1 group ( P<0.05). Compared with IR+ LV-GRSF1 group, the percentage of cerebral infarction volume was significantly increased, the count of viable neurons was decreased, the expression of p16, p21 and TNF-α mRNA in ischemic brain tissues was up-regulated, SOD and GSH contents were decreased, the MDA content was increased, the expression of GRSF1 and GPX4 was down-regulated, and the expression of ACSL4 and ferritin was up-regulated in IR+ LV-GRSF1+ RSL3 group ( P<0.05). Conclusions:GRSF1 is involved in the endogenous protective mechanism against cerebral I/R injury by up-regulating GPX4 expression, attenuating oxidative stress, and thus inhibiting ferroptosis in mice.

Relationship between cGAS-STING signaling pathway and ferritinophagy in early stage of cerebral ischemia-reperfusion in mice / 中华麻醉学杂志

Objective:To evaluate the relationship between the second messenger cyclic GMP-AMP (cGAS)-cyclic GMP-AMP receptor stimulator of interferon genes (STING) signaling pathway and ferritinophagy in the early stage of cerebral ischemia-reperfusion (I/R) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 21-25 g, were divided into 4 groups ( n=6 each) using a random number table method: sham group, cerebral I/R injury group (CIRI group), cerebral I/R injury + cGAS inhibitor group (CIRI + RU group), and cerebral I/R injury + cGAS inhibitor + overexpressed nuclear receptor coactivator 4 (NCOA4) group (MCAO + RU + LV-NCOA4 group). The model of cerebral I/R injury was developed using the middle cerebral artery occlusion (MCAO) in anesthetized animals.In CIRI+ RU group, cGAS inhibitor 5 mg/kg was intraperitoneally injected at 10 min before reperfusion.In CIRI+ RU+ LV-NCOA4 group, NCOA4-overexpressing lentivirus (1×10 9 TU/ml) 2 μl was injected into the ventricle at 7 days before MCAO, and the other operations were the same as those previously described in CIRI+ RU group.After 6 h of reperfusion, the neurological function deficits were assessed and scored, then the mice were sacrificed, and brains were removed for determination of the cerebral infarct size (by TTC method), MDA content (by TBA method), activity of SOD (by WST-1 method), and expression of cGAS, STING, NCOA4, ferritin, and microtubule-associated protein 1 light chain 3B (LC3B) (by Western blot). Results:Compared with Sham group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05). Compared with CIRI group, the neurological function deficit score and cerebral infarct size were significantly decreased, SOD activity was increased, MDA content was decreased, the expression of cGAS, STING, NCOA4 and LC3B was down-regulated, and the expression of ferritin was up-regulated in CIRI+ RU group ( P<0.05). Compared with CIRI+ RU group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05), and no significant change was found in the expression of cGAS and STING in CIRI+ RU+ LV-NCOA4 group ( P>0.05). Conclusions:The cGAS-STING signaling pathway can promote the over-activation of ferritinophagy, enhance oxidative stress, and thus induce early CIRI in mice.

Effect of SIRT3 overexpression on hypoxia-reoxygenation injury to hippocampal neurons of mice exposed to high glucose: relationship with SOD2 / 中华麻醉学杂志

Objective:To evaluate the effect of sirtuin 3 (SIRT3) overexpression on hypoxia-reoxygenation (H/R) injury to hippocampal neurons of mice exposed to high glucose and its relationship with SOD2.Methods:The normally cultured HT22 neurons at the logarithmic phase were selected and divided into 3 groups ( n=12 each) using a random number table method: high-glucose normoxia group (HG group), high glucose+ H/R group (HHR group) and high glucose+ H/R+ SIRT3 overexpression group (HHR+ SIRT3 group). To establish high glucose model, the neurons in 3 groups were cultured in high-glucose culture medium (glucose concentration of 50 mmol/L) for 8 h. In HHR and HHR+ SIRT3 groups, the cells were exposed to glucose-free and hypoxia for 6 h and then cultured in the high-glucose normoxic environment for 24 h to establish the high glucose and HR injury model.In HHR+ SIRT3 group, the neurons were transfected with SIRT3 overexpressed lentivirus.The cell viability was recorded by the cell counting kit-8 assay, reactive oxygen species (ROS) content was detected by flow cytometry, mitochondrial malonaldehyde (MDA) content, superoxide dismutase (SOD) activity, catalase (CAT) activity and adenosine triphosphate (ATP) content were determined by colorimetry, mitochondrial membrane potential (MMP) was detected by JC-1 probe, and the expression of nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), SIRT3, SOD2 and acetylated SOD2 (ac-SOD2) was detected by Western blot. Results:Compared with HG group, cell viability, SOD activity, CAT activity, ATP content, MMP, NRF1 and the expression of TFAM and SIRT3 were significantly decreased, and ROS content, MDA content and ac-SOD2/SOD2 ratio were increased in group HHR and group HHR+ SIRT3 ( P<0.05). Compared with HHR group, cell viability, SOD activity, CAT activity, ATP content, MMP, NRF1 and the expression of TFAM and SIRT3 were significantly increased, and ROS content, MDA content and ac-SOD2 /SOD2 ratio were decreased in HHR+ SIRT3 group ( P<0.05). Conclusion:SIRT3 overexpression can alleviate hypoxia-reoxygenation (H/R) injury to hippocampal neurons of mice incubated in high glucose medium, and the mechanism is related to activation of SOD2 deacetylation.

Role of autophagy in reduction of high glucose and hypoxia-reoxygenation injury to isolated cardiomyocytes by dexmedetomidine in rats / 中华麻醉学杂志

Objective:To evaluate the role of autophagy in reduction of high glucose and hypoxia-reoxygenation (HG+ H/R) injury to isolated cardiomyocytes by dexmedetomidine in rats.Methods:The normally cultured rat H9c2 cardiomyocytes at the logarithmic growth phase were seeded in 6-well plates at a density of 1×10 6 cells/ml and divided into 4 groups ( n=15 each) using a random number table method: control group (group C), group HG+ H/R, dexmedetomidine group (group DEX) and dexmedetomidine+ autophagy inhibitor 3-methylpurine group (group 3-MA). The cells were incubated in culture medium with 1% fetal bovine serum + 1% double antibody for 24 h when the cell density reached 50%.To establish HG+ H/R injury model, the cardiomyocytes were cultured in high-glucose culture medium (glucose concentration of 33 mmol/L) for 24 h, and then incubated in a 37 ℃ incubator (95% N 2+ 5%CO 2) for 4 h followed by reoxygenation (90%O 2+ 10%CO 2) for 2 h. Dexmedetomidine was added until the final concentration reached 5 μmol/L at 1 h before hypoxia in DEX and 3-MA groups, and 3-MA was added until the final concentration reached 5 μmol/L at 1 h of incubation with dexmedetomidine.At 2 h after reoxygenation, the cell viability was recorded by the cell counting kit-8 assay, lactate dehydrogenase (LDH) activity was detected by LDH kit, the expression of autophagy-related protein LC3, P62 and Beclin-1 was detected by Western Blot, the ratio of LC3Ⅱ/LC3Ⅰwas calculated, and the expression of P62 and Beclin-1 mRNA was detected by real-time polymerase chain reaction. Results:Compared with group C, the cell viability was significantly decreased in HG+ H/R, DEX and 3-MA groups, LDH activity in the supernatant was increased and expression of P62 was decreased in HG+ H/R and 3-MA groups, ratio of LC3Ⅱ/LC3Ⅰwas decreased in group 3-MA, and the expression of Beclin-1 was down-regulated in group HG+ H/R ( P<0.05). Compared with HG+ H/R group, LDH activity in the supernatant was significantly decreased, expression of Beclin-1, P62 and its mRNA was up-regulated, and ratio of LC3Ⅱ/LC3Ⅰwas increased in DEX group, and LDH activity in the supernatant was increased in group 3-MA ( P<0.05). Compared with DEX group, cell viability were decreased, LDH activity in the supernatant was increased, Beclin-1, P62 and its mRNA was down-regulated, and ratio of LC3Ⅱ/ LC3Ⅰwas decreased in group 3-MA ( P<0.05). Conclusion:Autophagy is involved in the reduction of HG+ H/R injury to isolated cardiomyocytes by dexmedetomidine in rats.

Role of Akt/GSK-3β signaling pathway in IL-4-induced reduction of cerebral ischemia-reperfusion injury in mice: the relationship with autophagy / 中华麻醉学杂志

Objective:To evaluate the role of B/glycogen synthase kinase-3β (Akt/GSK-3β) signaling pathway in interleukin-4 (IL-4)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in mice and the relationship with autophagy.Methods:Forty clean-grade healthy male Balb/c mice, aged 10-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), cerebral I/R group (group IR), IR plus IL-4 group, and IR plus IL-4 plus Akt inhibitor LY294002 group (IR+ IL-4+ LY group). Cerebral I/R was induced by 60 min middle cerebral artery occlusion followed by 24 reperfusion in anesthetized mice.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model in group IL-4.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model, and LY294002 15 nmol/kg was simultaneously injected via the tail vein in group IR+ IL-4+ LY.Neurological function was assessed and scored at 24 h of reperfusion, and then animals were sacrificed and brains removed for determination of cerebral infarct size (by TTC assay), cell apoptosis, autophagosome count (with transmission electron microscope), levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) (using colorimetric assay), phosphorylation of Akt and GSK-3β, expression of LC3 and Beclin-1 (by Western blot). The apoptosis index and LC3Ⅱ/LC3Ⅰ ratio were calculated. Results:Compared with Sham group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in the other three groups ( P<0.05). Compared with IR group, the neurological scores, cerebral infarct size and apoptosis index were significantly decreased, the SOD activity in brain tissues was increased, levels of MDA and ROS were decreased, phosphorylation of Akt and GSK-3β was increased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were decreased in IR+ IL-4 group ( P<0.05), and no significant change was found in the parameters mentioned above in IR+ IL-4+ LY group ( P>0.05). Compared with IR+ IL-4 group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in IR+ IL-4+ LY group ( P<0.05). Conclusion:IL-4 can inhibit cell autophagy through activating Akt/GSK-3β signaling pathway and thus reduces cerebral I/R injury in mice.

On the application of blended learning in normal human morphology course / 中华医学教育探索杂志

Objective:To explore the application effect of the blended learning based on Rain Classroom in normal human morphology course.Methods:A total of 118 undergraduates majoring in medical laboratory technology of Batch 2017 and Batch 2018 from Xinjiang Medical University were included in this study, and they were divided into the observation group (Batch 2018) and the control group (Batch 2017), with 59 students in each group, adopting the traditional teaching method and blended learning method based on Rain Classroom respectively. The mid-term, final and comprehensive scores of the two groups were compared by t test and chi-square test through SPSS 23.0. And the evaluation of teaching satisfaction of the observation group was obtained by the questionnaire survey. Results:The mid-term results [(74.02±8.71) vs. (62.00±8.97), t=-6.073, P<0.001], the final results [(83.21±7.73) vs. (70.44±11.43), t=-6.250, P<0.001], and the comprehensive results [(82.26±9.53) vs. (70.52±11.09), t=-6.012, P<0.001] of the observation group were significantly superior than those of the control group ( P<0.05). The excellence rate of the final results (23.72% vs. 3.45%, χ2=10.412, P=0.001) and comprehensive results (18.64% vs. 5.08%, χ2=5.187, P=0.023) in the observation group were significantly higher than those in the control group ( P<0.05). The results of the questionnaire survey showed that the observation group generally had a good satisfaction with the blended learning. Conclusion:The blended learning could make up for the deficiency of the traditional teaching methods, have preferable teaching effects, and get wide recognition from students.

The method of removing breathing baseline in pulse wave signal / 中国医疗器械杂志

A method which combines empirical mode decomposition with wavelet transform is employed to remove breathing baseline draft from pulse wave signal. First of all, this method decomposes pulse wave signal into several intrinsic mode functions and judges the intrinsic mode function which contains the information of breathing baseline draft. And then wavelet transform is used to decompose these intrinsic mode functions, and the detail coefficients representing breathing baseline draft are set into zero. At last, the signal is rebuilt. This can realize removing breathing baseline draft. A self-developed measurement device was used to obtain the pulse wave signal for validating, and AC-DC modulation ratio value was adopted to evaluate the effect. The results showed that this method could effectively remove breathing baseline draft from pulse wave signal.

A study of correlation between insulin resistance and the clinical pathology characters in chronic hepatitis B patients / 中国医师杂志

Objective To investigate the relationship between insulin resistance index (HOMA-IR) and the clinical pathology fea-tures in chronic hepatitis B(CHB) patients. Methods ALT and hepatitis B virus loading level and HOMA-IR were measured in 80 CHB pa-tients. Correlation analysis was performed with pathological features of liver tissue. Result Insulin resistance(IR) was much more obvious in patients with severer degree of liver inflammation and fibrosis . HOMA-IR and the insulin level of CHB patients in G3 grade(15.24±7.41,3.51 ± 1.91) and S3 stage (16.65±6. 46,3.79 ± 1.50) were higher than those in G2 grade (10.33 ± 6.28,2.25 ± 1.39) and S2 stage(9.06 ± 4.61,2.02 ± 1.11). The HOMA-IR was positive correlated with the level of age and ALT. Conclusion The development of IR in CHB patients was correlated with age and the degree of liver inflammation and fibrosis.

Clinical observation on Herba Houttuyniae Injection in treating senile pneumonia / 国际中医中药杂志

Objective To observe the clinical effect of Herba Houttuyniae Injection in treating senile pneumonia and to explore its possible mechanism. Methods Sixty-five patients were randomly divided into two groups. The control group was treated with conventional western therapy (CWT), while the treatment group was treated with CWT plus Herba Houttuyniae Injection. The changes of serum cytokines, IL-2 and TNF-?, before and after treatment were observed. Results the total effective rate of treatment group is 94.3%, while that of the control group is 83.3%. The difference of the total effective rate between the two groups was significant(P

Determination of insulin resistance index and its clinical significance in chronic liver disease related to hepatitis B virus / 解放军医学杂志

Objective To investigate the role of insulin resistance index (HOMA-IR) in premonishing hepatogenous diabetes subsequent to chronic liver disease related to hepatitis B virus. Methods Fasting plasma glucose (FPG) and fasting serum insulin (FINS) were measured in 176 patients, including 80 patients with chronic hepatitis B (CHB), 66 patients with liver cirrhosis (LC) and 30 patients with chronic severe hepatitis B (CSHB). In addition 30 healthy subjects served as control (group HC). None of the subjects in the study had a history of diabetes. FPG and FINS were measured in all the subjects to exclude impaired glucose tolerance and diabetes. Then HOMA-IR was calculated. Results Among the HC and CHB, LC and CSHB patients, both FINS and HOMA-IR levels gradually elevated. FINS and HOMA-IR in CHB patients (10.58?6.48, 2.35?1.55) were significantly higher than those in group HC (8.06?2.14,1.68?1.53), while lower than those in LC (12.73?5.88, 2.91?1.41) and CSHB (18.30?14.17, 4.35?3.17) patients (P0.05), while the values of FINS and HOMA-IR were significantly higher in severe patients than those in moderate patients (P0.05), while the values of FPG were significantly higher in LC and CSHB patients than those in group HC (P