Role of ROS in attenuation of hypoxia-reoxygenation injury in rat cardiomyocytes by pinacidil postconditioning: relationship with Nrf2-ARE signaling pathway / 中华麻醉学杂志

Objective:To evaluate the role of reactive oxygen species (ROS) in attenuation of hypoxia-reoxygenation (H/R) injury in rat cardiomyocytes by pinacidil postconditioning and the relationship with nuclear factor erythrid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway.Methods:Adult rat cardiomyocytes were isolated and cultured and then divided into 4 groups ( n=20 each) by a random number table method: control group (group C), H/R group, pinacidil postconditioning group (group P) and reactive oxygen scavenger N-(2-mercaptopropionyl)-glycine(MPG)+ pinacidil postconditioning group (group MPG+ P). Group C was continuously exposed to 95%O 2+ 5%CO 2 in an incubator at 37 ℃ for 105 min. The cells were exposed to 5%CO 2+ 1%O 2+ 94%N 2 in an incubator at 37 ℃ for 45 min followed by reoxygenation for 60 min to prepare H/R injury model. The cells were exposed to hypoxia for 45 min and then treated with pinacidil 50 μmol/L for 5 min followed by reoxygenation for 60 min in group P. The cells were exposed to hypoxia for 45 min, treated with MPG 2 mmol/L for 10 min, and then treated with pinacidil for 5 min followed by reoxygenation for 60 min in group MPG+ P. The content of Ca 2+ and activity of Nrf2 in cardiomyocytes were measured at the end of reoxygenation. The ultrastructure of cardiomyocytes was observed, and mitochondrial ultrastructure was evaluated using mitochondrial Flameng score. The expression of Nrf2, superoxide dismutase (SOD1), quinone oxidoreductase 1 (NQO1), and heme oxygenase 1 (HO-1) protein and mRNA was detected using Western blot and real-time polymerase chain reaction. Results:Compared with group C, the Ca 2+ content, Nrf2 activity and mitochondrial Flameng score were significantly increased, the expression of Nrf2, SOD1, NQO1 and HO-1 protein and mRNA was down-regulated ( P<0.05), and the damage to the ultrastructure of cardiomyocytes was aggravated in group H/R. Compared with H/R group, the Ca 2+ content and mitochondrial Flameng score were significantly decreased, the Nrf2 activity was increased, the expression of Nrf2, SOD1, NQO1 and HO-1 protein and mRNA was up-regulated ( P<0.05), and the damage to the ultrastructure of cardiomyocytes was attenuated in P group. Compared with P group, the Ca 2+ content and mitochondrial Flameng score were significantly increased, the Nrf2 activity was decreased, the expression of Nrf2, SOD1, NQO1 and HO-1 protein and mRNA was down-regulated ( P<0.05), and the damage to the ultrastructure of cardiomyocytes was aggravated in MPG+ P group. Conclusions:ROS is involved in attenuation of H/R injury by pinacidil postconditioning, which is associated with activation of the Nrf2-ARE signaling pathway in rat cardiomyocytes.

Relationship between lung injury caused by cardiopulmonary bypass and acetyltransferase p300 in rats / 中华麻醉学杂志

Objective:To evaluate the relationship between lung injury induced by cardiopulmonary bypass (CPB) and acetyltransferase p300 (p300) in rats.Methods:Eighteen SPF healthy adult male Sprague-Dawley rats, aged 12-16 weeks, weighing 350-450 g, were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (S group), CPB group, and CPB+ left lung ischemia-reperfusion (I/R) group (CPB+ IR group). CPB group was connected to CPB pipeline for cardiopulmonary bypass. The lung I/R injury model was prepared by clamping the left lung hilum for 45 min followed by opening during CPB, 30 min later CPB was terminated, and mechanical ventilation was continuously performed for 1.5 h before ending the experiment in CPB+ IR group. Arterial blood gas analysis was performed and oxygenation index (OI) and respiratory index (RI) were calculated before CPB, at 10 min after opening the lung hilum, and immediately after the end of experiment. The bronchoalveolar lavage fluid (BALF) and left lung tissues were collected immediately after the end of experiment for determination of the concentrations of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in BALF and total protein in BALF and concentrations of IL-17 in lung tissues (by enzyme-linked immunosorbent assay), expression of p300, phosphorylated p300 (p-p300), and acetylated histone H3 (AC-H3) in lung tissues (by Western blot) and expression of p-p300 (using immunohistochemical staining) and for microscopic examination of the pathological changes of lung tissues (under the light microscope) which were scored. Results:Compared with S group, OI was significantly decreased and RI was increased at 10 min after opening the lung hilum and immediately after the end of experiment, the lung injury score and levels of IL-6, TNF-α and total protein in BALF and IL-17 in lung tissues were increased, and the expression of p300, p-p300 and AC-H3 was up-regulated in CPB and CPB+ IR groups ( P<0.05). Compared with CPB group, OI was significantly decreased and RI was increased at 10 min after opening the lung hilum and immediately after the end of experiment, the lung injury score and levels of IL-6, TNF-α and total protein in BALF and IL-17 in lung tissues were increased, and the expression of p300, p-p300 and AC-H3 was up-regulated in CPB+ IR group ( P<0.05). Conclusions:The mechanism by which CPB induces lung injury may be related to up-regulation of the expression of p300 and enhancement of activity of p300 in lung tissues and increased release of inflammatory factors in rats.

Preliminary study on the regulation of acute myeloid leukemia by FLT3 gene expression / 中华医学遗传学杂志

OBJECTIVE@#To assess the influence of FLT3 expression on the prognosis of patients with acute myeloid leukemia (AML) by cell experiment and clinical data analysis.@*METHODS@#Models for FLT3 over-expression and interference-expression in AML cells were constructed. The level of BAK gene expression and its protein product was determined, along with the proliferation and apoptosis of leukemia cells. FLT3 gene expression and FLT3-ITD variant were determined among patients with newly diagnosed AML.@*RESULTS@#Compared with the interference-expression group, the level of BAK gene expression and its protein in FLT3 over-expression AML cells was significantly lower (P < 0.001), which also showed significantly faster proliferation (P < 0.001) and lower rate of apoptosis (P < 0.001). The expression level of FLT3 gene among patients with newly diagnosed AML was also significantly higher compared with the healthy controls (P < 0.001). The FLT3 gene expression of FLT3-ITD positive AML patients was higher than that of FLT3-WT patients (P = 0.002). Survival analysis showed that AML patients with high FLT3 expression in the medium-risk group had a lower complete remission rate and overall survival rate compared with those with a low FLT3 expression (P < 0.001).@*CONCLUSION@#Over-expression of FLT3 may influence the course of AML by promoting the proliferation of leukemia cells and inhibiting their apoptosis, which in turn may affect the prognosis of patients and serve as a negative prognostic factor for AML.

Epileptogenicity of different types of cortical tubers in patients with tuberous sclerosis complex / 中华实用儿科临床杂志

Objective:To record stereoelectroencephalography (SEEG) data and to induce cortical electrical stimulation in children with tuberous sclerosis complex (TSC), thus exploring the epileptogenicity of different types of cortical tubers.Methods:The SEEG recording and cortical electrical stimulation data of 50 children with TSC who underwent preoperative evaluation for drug-resistant epilepsy at Epilepsy Center, Tsinghua University Yuquan Hospital from November 2016 to September 2022 were retrospectively analyzed, involving 27 boys and 23 girls with the age of (5.5±3.4) years.According to the results of 3.0T magnetic resonance imaging (3T-MRI) and computed tomography(CT), cortical tubers were classified.The incidences of electroclinical seizures, electrical seizures and seizures induced by cortical electrical stimulation in different types of tubers recorded by SEEG were analyzed, and the differences in the proportion of the above seizures among different types of tubers were compared using the Fisher′ s exact test. Results:A total of 303 cortical tubers were explored using SEEG in 50 patients.The tubers were divided into 6 types, including Type A, B, C, D and E, and focal cortical dysplasia like (FCD-like) type, among which Type E was for the first time proposed in the world.Among these explored tubers, 7 tubers had electrical seizures, and 57 tubers had electroclinical seizures.A total of 64 tubers (21.1%) were epileptogenic.The incidence of epileptogenic tubers in Type A-E and FCD-like type were 3.6%, 1.4%, 19.0%, 77.8%, 77.5%, and 90.0%, respectively. Fisher′ s exact test and Bonferroni correction were performed for pairwise comparisons( P<0.003). There was no significant difference in the incidence of epileptogenic tubes among Type A, B and C. There was significant difference in the incidence of epileptogenic tubes between Type A-C with Type D, Type E and FCD-like type, respectively.There was no significant difference in the incidence of epileptogenic tubes between Type D, Type E and FCD-like like.Electrical stimulation-induced seizures occurred in 36 cortical tubers (11.9%). The positive rate of electrical stimulation seizures in Type A-E and FCD-like type were 0.7%, 1.4%, 4.8%, 44.4%, 45.0%, and 70.0%, respectively.There was significant difference in the positive rate of electrical stimulation seizures between Type A-B and Type D, Type E and FCD-like type, respectively, so as that between Type C versus Type E and FCD-like type.No significant difference in the positive rate of electrical stimulation seizures was found between other pairwise comparisons. Conclusions:This study proposed a new classification of cortical tubers in TSC patients, and Type E is proposed for the first time in the world.SEEG records confirmed great differences in epileptogenicity indifferent types of cortical tubers.Type D, Type E and FCD-like type have higher epileptogenicity, which is of great value for the preoperative evaluation of TSC epilepsy surgery and the placement strategy of SEEG electrodes.

Study on prediction of potential targets and mechanism of Zeqi Decoction in the treatment of lung adenocarcinoma / 国际中医中药杂志

Objective:To explore the potential molecular targets and mechanism of Zeqi Decoction in the treatment of lung adenocarcinoma (LUAD) through bioinformatics and cell experiment.Methods:The active components of Zeqi Decoction were collected based on TCMSP database and literature search. Then R software was used to screen differentially expressed genes in LUAD from TCGA and GEO databases. The co-expression module was obtained through weighted gene co-expression network analysis (WGCNA), and the potential targets were obtained after matching and mapping with targets of Zeqi Decoction. Enrichment analysis of GO function and KEGG pathway of targets was conducted. The results were experimentally verified. The lung adenocarcinoma cell lines A549 and H1299 were divided into blank control group and Zeqi Decoction group according to random number table. The inhibition rate of cell proliferation was detected by cell proliferation test (CCK-8); the expression of leukocyte differentiation antigen 36 (CD36) in A549 and H1299 cells was detected by Western blot; the levels of low density lipoprotein receptor (LDLR) and IL-6 were detected by ELISA.Results:Totally 157 anti-lung adenocarcinoma active components and 18 potential targets were obtained, mainly including CD36, IL6, LDLR, etc. The main target of Zeqi Decoction in the treatment of lung adenocarcinoma was lipid metabolism. The results showed that Zeqi Decoction could effectively inhibit the activity of A549 and H1299 cells and the levels of CD36, LDLR and IL-6.Conclusion:Zeqi Decoction can inhibit the inflammatory response by down-regulating the protein expressions of CD36 and LDLR, thereby slowing the proliferation of cells.

The characteristics of stereoelectroencephalography in drug-resistant epileptic spasms / 中华实用儿科临床杂志

Objective:To analyze the characteristics of stereoelectroencephalography (SEEG) in children with drug-resistant epileptic spasms (ES), and to explore the surgical strategy of children with spastic seizure under the guidance of SEEG.Methods:The clinical data of 156 children with ES who were preoperatively evaluated in the Department of Neurosurgery Ward 3, Tsinghua University Yuquan Hospital from January 2014 to December 2021 were retrospectively reviewed.All children were evaluated in the second stage of stereotactic electrode placement after a non-invasive preoperative evaluation.The characteristics of intracranial EEG, surgical strategy and prognosis were analyzed.Results:A total of 19 eligible children were included, involving 13 boys and 6 girls.The age of first onset and surgical age of them ranged 1 month to 4 years, and 2 years to 13 years, respectively.The SEEG was divided into 3 types in children with ES at the onset.Five children were SEEG type A, presenting with the focal seizure discharges at the beginning and a gradual propagation to widespread fast-wave bursts.Ten children were SEEG type B, presenting a focal leading spike followed by diffused fast-wave bursts.Four children were SEEG type C, presenting a diffuse fast wave rhythm onset.Although some electrode discharges appeared slightly " leading", they covered more than one brain region.After focal resection or thermocoagulation, 13/19 patients did not have the onset of seizures, and 5/19 and 8/19 were graded as SEEG type A, and B, respectively.During the intermittent period of SEEG attacks in children with SEEG type A and B, a significant phenomenon of focal epileptic discharge consistent with the onset of the attack was observed, and surgical removal of these areas effectively controlled spastic seizures.Conclusions:Epileptic spasms may be triggered by a focal neocortical discharge.Intracranial EEG showed that the focal seizure onset evolves into spasm or a focal " leading spike" is a good indicator of surgical prognosis.

Role of succinate dehydrogenase in hypoxic postconditioning-induced reduction of hypoxia-reoxygenation injury in myocardial cells of rats and the relationship with mito-K ATP / 中华麻醉学杂志

Objective:To evaluate the role of succinate dehydrogenase (SDH) in hypoxic postconditioning (HPC)-induced reduction of hypoxia-reoxygenation (H/R) injury in myocardial cells of rats and the relationship with mitochondrial ATP-sensitive potassium channels (mito-K ATP). Methods:Myocardial cells isolated from adult male Sprague-Dawley rats were cultured for 48 h and then divided into 7 groups ( n=24 each) using a random number table method: blank control group (Nor group), H/R group, SDHA-siRNA adenovirus+ H/R group (siRNA+ H/R group), HPC group, SDHA-siRNA adenovirus+ HPC group (siRNA+ HPC group), 5-HD+ HPC group, and SDHA-siRNA adenovirus+ 5-HD+ HPC group (siRNA+ 5-HD+ HPC group). Nor group was continuously cultured for 195 min under normoxic conditions. The H/R injury model was prepared by exposing the cells to hypoxia for 45 min in 5% CO 2 + 1% O 2 + 94% N 2, followed by reoxygenation for 150 min. The HPC method involved three cycles of 5 min reoxygenation/5 min hypoxia at the end of 45 min ischemia before 120 min reoxygenation. The mito-K ATP blocker 5-HD administration method involved adding 5-HD at a final concentration of 100 μmol/L at 30 min of hypoxia. The myocardial cells in each siRNA group were successfully transfected with SDHA-siRNA adenovirus to silence SDHA expression. The cell viability, calcium ion level, SDH activity, ATP content, degree of mitochondrial permeability transition pore (mPTP) opening, and mitochondrial membrane potential (MMP) were measured at the end of reoxygenation. Results:Compared with Nor group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, level of calcium ion and activity of SDH were increased in H/R group ( P<0.05). Compared with H/R group, the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ H/R group and HPC group ( P<0.05). Compared with HPC group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, calcium ion level and SDH activity were increased in 5-HD+ HPC group ( P<0.05), and the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ HPC group ( P<0.05). Compared with siRNA+ HPC group, the cell viability, ATP content and MMP were significantly decreased, the opening degree of mPTP and calcium ion level were increased ( P<0.05), and no significant change was found in the SDH activity in siRNA+ 5-HD+ HPC group ( P>0.05). Compared with 5-HD+ HPC group, the SDH activity was significantly decreased, and no significant change was found in the other parameters in siRNA+ 5-HD+ HPC group ( P>0.05). Conclusions:HPC alleviates H/R injury probably by reducing SDH activity and opening mito-K ATP in myocardial cells of rats.

Role of reactive oxygen species in hypoxia postconditioning-induced activation of Nrf2/ARE signaling pathway in rat cardiomyocytes / 中华麻醉学杂志

Objective:To evaluate the role of reactive oxygen species (ROS) in hypoxia postconditioning-induced activation of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway in rat cardiomyocytes.Methods:Primary cardiomyocytes of adult rats were isolated and cultured and divided into 4 groups ( n=20 each) using a random number table method: normal group (group N), hypoxia-reoxygenation group (group HR), hypoxia postconditioning group (group HPO) and hypoxia postconditioning plus an ROS scavenger N-(2-Amidinopropionyl)-glycine (MPG) group (group HPO+ MPG). Cells were exposed to hypoxia for 45 min followed by 60 min reoxygenation to develop the cardiomyocyte hypoxia-reoxygenation injury model.In HPO group, cells were subjected to 3 cycles of 5-min hypoxia/5-min reoxygenation after 45 min hypoxia, followed by reoxygenation for 60 min.In HPO+ MPG group, MPG (final concentration 2 mmol/L) was added at 35 min of hypoxia, cells were subjected to hypoxia for 10 min, and the other treatments were similar to those previously described in group HPO.At the end of reoxygenation, the intracellular calcium level and Nrf2 activity were measured, the ultrastructure of cardiomyocytes was observed, and the Flameng score of mitochondria was assessed, and the expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (NQO1), superoxide dismutase 1 (SOD1) and heme oxygenase-1 (HO-1) protein and mRNA was detected using real-time polymerase chain reaction and Western blot. Results:Compared with group N, the intracellular free Ca 2+ level, Nrf2 activity and Flameng score were significantly increased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was down-regulated in group HR ( P<0.05). Compared with group HR, the intracellular free Ca 2+ level and Flameng score were significantly decreased, the Nrf2 activity was increased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was up-regulated in group HPO ( P<0.05). Compared with group HPO, the intracellular free Ca 2+ level and Flameng score were significantly increased, the Nrf2 activity was decreased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was down-regulated in group HPO+ MPG ( P<0.05). Conclusions:The mechanism by which hypoxia postconditioning activates the Nrf2/ARE signaling pathway in rat cardiomyocytes may be related to ROS.

Mitochondrial mechanism of diabetic mellitus-caused abolition of cardioprotection induced by ischemia postconditioning in rats: succinate dehydrogenase / 中华麻醉学杂志

Objective:To evaluate the relationship between the mitochondrial mechanism of diabetic mellitus-caused abolition of cardioprotection induced by ischemia postconditioning (IPO) and succinate dehydrogenase (SDH) in rats.Methods:Thirty-six SPF male non-diabetic Sprague-Dawley rats, aged 16-20 weeks, weighing about 300 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (ND+ Sham group), ischemia-reperfusion (I/R) group (ND+ I/R group) and IPO group (ND+ IPO group). Seventy-two rats with diabetes mellitus were divided into 6 groups ( n=12 each) using a random number table method: sham operation group (DM+ Sham group), I/R group (DM+ I/R group), DM+ IPO group, sham operation+ dimethyl malonate group (group DM+ Sham+ Dme), I/R+ dimethyl malonate group (group DM+ I/R + Dme) and IPO+ dimethyl malonate group (group DM+ IPO+ Dme). The model of cardiopulmonary bypass (CPB) was established, and the model of total I/R injury was induced by ligating the ascending aorta for 30 min followed by 60 min of reperfusion.The animals underwent 3 cycles of 30-s reperfusion followed by 30-s ischemia starting from the onset of reperfusion in each IPO group.In each Dme group, dimethyl malonate was infused through the tail vein at a rate of 4 mg· kg -1·min -1 for 40 min starting from the beginning of CPB.At the end of reperfusion, the myocardial tissues were taken for measurement of mitochondrial respiratory control ratio (RCR) (by the Lufthansa electrode method), mitochondrial membrane potential (MMP) (by the JC-1 method) and the opening of mitochondrial permeability transition pore (mPTP) (by absorptiometry) and for determination of the activity of reactive oxygen species (ROS) (with the fluorescent probe), succinate dehydrogenase (SDH) (using spectrophotometric method) and the contents of succinic acid and fumarate. Results:Compared with ND+ Sham group, the activities of SDH and ROS, opening of mPTP and content of fumarate were significantly increased, and MMP, RCR and succinic acid content were decreased in ND+ I/R ( P<0.05). Compared with group ND+ I/R, the activities of SDH and ROS, opening of mPTP and content of fumarate were significantly decreased, and MMP, RCR and succinic acid content were increased in ND+ IPO ( P<0.05). Compared with group DM+ Sham, the activities of SDH and ROS, opening of mPTP and content of fumarate were significantly increased, and MMP, RCR and succinic acid content were decreased in group DM+ I/R ( P<0.05). Compared with group DM+ I/R, no significant change was found in the parameters mentioned above in group DM+ IPO ( P>0.05). Compared with group DM+ IPO, the activities of SDH and ROS, opening of mPTP and content of fumarate were significantly decreased, and MMP, RCR and succinic acid content were increased in group DM+ IPO+ Dme ( P<0.05). Compared with group DM+ I/R+ Dme, the activities of SDH and ROS, opening of mPTP and content of fumarate were significantly decreased, and MMP, RCR and succinic acid content were increased in group DM+ IPO+ Dme ( P<0.05). Conclusion:The mitochondrial mechanism of diabetic mellitus-caused abolition of cardioprotection induced by IPO may be related to the enhancement of SDH activity in rats.

Orlistat induces ferroptosis-like cell death of lung cancer cells / 医学前沿

Aberrant de novo lipid synthesis is involved in the progression and treatment resistance of many types of cancers, including lung cancer; however, targeting the lipogenetic pathways for cancer therapy remains an unmet clinical need. In this study, we tested the anticancer activity of orlistat, an FDA-approved anti-obesity drug, in human and mouse cancer cells in vitro and in vivo, and we found that orlistat, as a single agent, inhibited the proliferation and viabilities of lung cancer cells and induced ferroptosis-like cell death in vitro. Mechanistically, we found that orlistat reduced the expression of GPX4, a central ferroptosis regulator, and induced lipid peroxidation. In addition, we systemically analyzed the genome-wide gene expression changes affected by orlistat treatment using RNA-seq and identified FAF2, a molecule regulating the lipid droplet homeostasis, as a novel target of orlistat. Moreover, in a mouse xenograft model, orlistat significantly inhibited tumor growth and reduced the tumor volumes compared with vehicle control (P < 0.05). Our study showed a novel mechanism of the anticancer activity of orlistat and provided the rationale for repurposing this drug for the treatment of lung cancer and other types of cancer.

EGFR tyrosine kinase inhibitor HS-10296 induces autophagy and apoptosis in triplenegative breast cancer MDA-MB-231 cells / 浙江大学学报·医学版

OBJECTIVE@#To investigate the inhibitory effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI) HS-10296 on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells and explore the possible molecular mechanism.@*METHODS@#MDA-MB-231 cells were treated with HS-10296 for 24, 48, or 72 h, and CCK-8 assay was used to assess the changes in the cell viability. The inhibitory effect of HS-10296 on cell proliferation was determined by clonogenic assay. JC-1 and flow cytometry were employed for analyzing the cell apoptosis, and the ultrastructure of the cells was observed under electron microscope. After pretreatment with autophagy inhibitor chloroquine (CQ), MDA-MB-231 cells were divided into control group, CQ treatment group, HS-10296 (4 and 6 μmol/L) treatment groups and combined treatment groups, and the sensitivity of the treated cells to HS-10296 was determined using CCK-8 assay. The effects of HS-10296 on EGFR pathway and apoptosis- and autophagy-related proteins in MDA-MB-231 cells were investigated using Western blotting.@*RESULTS@#HS-10296 significantly inhibited the proliferation of MDA-MB-231 cells with IC values at 24, 48 and 72 h of 8.393, 2.777 and 2.016 μmol/L, respectively. JC-1 and flow cytometry showed that HS-10296 induced obvious apoptosis of MDA-MB-231 cells, which showed an apoptosis rate of (21.63 ± 2.97)% following treatment with 8 μmol/L HS-10296. Autophagy vesicles were observed in the cells treated with HS-10296 under electron microscope. In MDA-MB-231 cells pretreated with CQ, inhibition of autophagy significantly enhanced HS-10296-induced cell death. Western blotting showed that the apoptosis-related protein caspase-3 was activated after HS-10296 treatment to cut its substrate PARP. The expression of autophagy-related protein light chain 3B (LC3B) was significantly enhanced after HS-10296 treatment ( < 0.01), which also resulted in inhibited phosphorylation of EGFR and AKT proteins in the cells.@*CONCLUSIONS@#HS-10296 can inhibit the proliferation and induce autophagy and apoptosis of MDA-MB-231 cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

EGFR tyrosine kinase inhibitor HS-10296 induces autophagy and apoptosis in triplenegative breast cancer MDA-MB-231 cells / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI) HS-10296 on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells and explore the possible molecular mechanism.@*METHODS@#MDA-MB-231 cells were treated with HS-10296 for 24, 48, or 72 h, and CCK-8 assay was used to assess the changes in the cell viability. The inhibitory effect of HS-10296 on cell proliferation was determined by clonogenic assay. JC-1 and flow cytometry were employed for analyzing the cell apoptosis, and the ultrastructure of the cells was observed under electron microscope. After pretreatment with autophagy inhibitor chloroquine (CQ), MDA-MB-231 cells were divided into control group, CQ treatment group, HS-10296 (4 and 6 μmol/L) treatment groups and combined treatment groups, and the sensitivity of the treated cells to HS-10296 was determined using CCK-8 assay. The effects of HS-10296 on EGFR pathway and apoptosis- and autophagy-related proteins in MDA-MB-231 cells were investigated using Western blotting.@*RESULTS@#HS-10296 significantly inhibited the proliferation of MDA-MB-231 cells with IC values at 24, 48 and 72 h of 8.393, 2.777 and 2.016 μmol/L, respectively. JC-1 and flow cytometry showed that HS-10296 induced obvious apoptosis of MDA-MB-231 cells, which showed an apoptosis rate of (21.63 ± 2.97)% following treatment with 8 μmol/L HS-10296. Autophagy vesicles were observed in the cells treated with HS-10296 under electron microscope. In MDA-MB-231 cells pretreated with CQ, inhibition of autophagy significantly enhanced HS-10296-induced cell death. Western blotting showed that the apoptosis-related protein caspase-3 was activated after HS-10296 treatment to cut its substrate PARP. The expression of autophagy-related protein light chain 3B (LC3B) was significantly enhanced after HS-10296 treatment ( < 0.01), which also resulted in inhibited phosphorylation of EGFR and AKT proteins in the cells.@*CONCLUSIONS@#HS-10296 can inhibit the proliferation and induce autophagy and apoptosis of MDA-MB-231 cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

Clinico-pathological investigations of a distinct variant of focal cortical dysplasia characterized by neuronal loss of layer four / 中华神经科杂志

Objective To investigate the clinical pathologic features of a distinct variant of focal cortical dysplasia (FCD) characterized by neuronal loss of layer four.Methods Between 2005 and 2017,approximately 3 000 surgeries were performed for the treatment of intractable epilepsy at Xuanwu Hospital,Capital Medical University and Yuquan Hospital,Tsinghua University.Retrospective analysis of clinic-pathological data of patients with epilepsy surgery was made and histological manifestations of neuronal loss of cortical layer four were included in this study.Results In this cohort,25 patients (22 males and three females) were identified with early onset pharmaco-resistant epilepsy and regionally circumscribed neuronal loss of cortical layer four in surgical specimens from the occipital lobe.Histologically,except for neuronal loss in cortical layer four in all cases,glial scar lesions were found in some patients.Thus the histology of those cases can be subdivided into two groups:group A (13 cases):neuronal loss of cortical layer four without glial scar lesions;and group B (12 cases):neuronal loss of cortical layer four with glial scar lesions.Due to the prominent horizontal disorganization of cortical layering and lack of any other microscopically visible principle lesion,group A should be classified hitherto as FCD International League Against Epilepsy (ILAE) type Ⅰ b,however,group B with scar lesions and cortical dysplasia around the main leision,should be classified as FCD ILAE type Ⅲd.This retrospective analysis of clinical histories revealed a perinatal distress in 20 patients (80％),suggesting an acquired pathomechanism.Magnetic resonance imaging revealed abnormal signals in the occipital lobe in all patients,and signal changes suggestive of encephalomalacia were found in 18 patients.Surgical treatment achieved favorable seizure control (Engel class Ⅰ and Ⅱ) in 18 patients (75％ among 24 available follow up).Comparion of the two groups with age at epilepsy onset (group A:5.00±2.76,group B:5.01±3.78),the proportion of perinatal distress (group A:11/13,group B:9/12) and the follow-up results (favorable seizure control of the two groups was 9/13,9/11 respectively) showed that there was no statistically significant difference between the two groups.Conculsion Neuronal loss of cortical layer four in the occipital lobe should be classified as a distinct variant of FCD ILAE type Ⅲd.

Role of mito-KATP channel in pinacidil postconditioning-induced reduction of myocardial ischemia-reperfusion injury in rats: an in vitro experiment / 中华麻醉学杂志

Objective To evaluate the role of mitochondrial KATP (mito-KATP) channel in pinacidil postconditioning-induced reduction of myocardial ischemia-reperfusion (I/R) injury in rats.Methods SPF healthy male Sprague-Dawley rats,aged 16-20 weeks,weighing 250-300 g,were anesthetized with pentobarbital.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 36.5-37.5 ℃.Thirty-two Langendorff-perfused hearts were divided into 4 groups (n =8 each) using a random number table:control group (group C),group I/R,pinacidil postconditioning group (group P) and 5-hydroxy decanoic acid plus pinacidil postconditioning group (group 5-HD+P).Myocardial ischemia was induced by interrupting perfusion for 40 min followed by 60 min reperfusion.Immediately after onset of reperfusion,hearts were perfused with K-H solution containing 50 μmol/L pinacidil for 2 min and then with K-H solution for 58 min in group P,hearts were perfused with K-H solution containing 100 μmol/L 5-HD for 5 min,with K-H solution containing 50 μmol/L pinacidil for 2 min and then with K-H solution for 53 min in group 5-HD+P.The heart rate (HR),left ventricular developed pressure (LVDP),left ventricular end-diastolic pressure (LVEDP) and the maximum rate of increase in left ventricular pressure (+dp/dtmax) were recorded at 20 min of equilibration (T1) and at the end of reperfusion (T2).Myocardial tissues were obtained at T2 for determination of myocardial infarct size and for examination of myocardial ultrastructure and Flameng scoring of the mitochondria was performed.Results Compared with group C,the HR,LVDP and +dp/dtmax were significantly decreased,and the LVEDP,myocardial infarct size and mitochondrial Flameng score were increased at T2 in group I/R (P＜0.05).Compared with group I/R,the HR,LVDP and +dp/dtmax were significantly increased and the LVEDP,myocardial infarct size and mitochondrial Flameng score were decreased at T2 (P＜0.05),and the pathological changes of myocardium were significantly attenuated in group P,and no significant change was found in the parameters mentioned above in group 5-HD+P (P＞0.05).Compared with group P,the HR,LVDP and + dp/dtmax were significantly decreased and the LVEDP,myocardial infarct size and mitochondrial Flameng score were increased at T2 (P＜O.05),and the pathological changes of myocardium were accentuated in group 5-HD+P.Conclusion The whole mechanism by which pinacidil postconditioning reduces myocardial I/R injury is related to promoting opening of mito-KATP channel in rats.

Analysis of pesticide poisoning in Ji'nan during 2012-2016 / 中华劳动卫生职业病杂志

Objective@#To study the current situation and distribution characteristics of pesticide poisoning in Ji'nan area, and to provide the basis for formulating the policy of scientific prevention and control of pesticide poisoning.@*Methods@#The cases of pesticide poisoning from 2012 to 2016 were collected from medical institutions in Ji'nan, and the data was subjected to statistical analysis.@*Results@#From 2012 to 2016 in Ji'nan reported a total of 2 237 cases of pesticide poisoning, non productive pesticide poisoning cases (72.78%, 2 149/2 237) and mortality (17.73%, 381/2 149) was significantly higher than that of productive pesticide poisoning. The average age is 46.78±18.57. The highest mortality rate of pesticide poisoning is more than 70 age group of the non productive pesticide poisoning. Organophosphorus pesticides (67.68%, 1 514/2 237) are the main pesticides causing poisoning, followed by herbicide (23.74%, 531/2 237). The highest mortality rate of pesticide was Paraquat (36.45%, 160/439), the second is the dichlorvos (19.19%, 170/886).@*Conclusion@#Pesticide poisoning is a public health problem and social problem which is harmful to the health of the residents in Ji'nan. It is necessary to strengthen the control and management of high toxic pesticides. Pay attention to the psychological intervention of elderly people to reduce the incidence of pesticide poisoning.

Research progress of treatments on colorectal cancer in elderly patients / 中华老年医学杂志

The patients aged ≥ 60 years account for 70％ of Chinese population with colorectal cancer.However,with the increase of age,the treatment methods such as surgery,radiotherapy,chemotherapy become inappropriate and impeding factors for the self-care of the elderly.Several clinical trials show that only a part of the elderly,who are in good health condition,can get the clinical benefits from these therapies.Therefore,this review focused on the comprehensive treatments for elderly patients with colorectal cancer,then reached the best individualized treatment,so as to improve the survival and quality of life.

Explore the occurrence and development of β cell dysfunction and insulin resistance according to the stratification on normal glucose tolerance / 中华内分泌代谢杂志

After the stratification of the normal glucose tolerance, the changes of insulin resistance and βcell function in the development of type 2 diabetes mellitus were investigated. A retrospective analysis on data of 275 cases with oral glucose insulin releasing tests. The area under the insulin curve (AUCINS ) 108. 43 mU/ L was taken as the critical value of diagnosis. Normal glucose tolerance subjects were divided into the NGT-a group(AUCINS<108. 43 mU/ L) and the NGT-b group(AUCINS≥108. 43 mU/ L). The plasma glucose, insulin, insulin sensitivity, and β cell function were compared among the 4 groups: NGT-a group (n=96), NGT-b group (n=49), prediabetes group (n=71), and type 2 diabetes mellitus group ( n = 59). Among the fasting insulin, 2 h insulin, AUCINS , early-phase insulin secretion index(△I30 / △G30), the ratio of total insulin area under curve, and total glucose area under curve, disposition index, homeostasis model assessment for insulin resistance, and Matsuda insulin sensitivity index, the relationship as follows: NGT-b group>prediabetes group>NGT-a group>type 2 diabetes mellitus group. The NGT-b group was always the highest, prediabetes group was lower, NGT-a group and type 2 diabetes mellitus group were the lowest, there were significant differences (all P<0. 05). Making the NGT-a group as the basic state, in the NGT-b group, β cell function has begun to appear compensation and insulin resistance, and β cell function compensation reached the peak, the β cell function in the prediabetes group was beginning to compensate for the deficiency, the function of β cell in type 2 diabetes mellitus group decreased further. These findings suggest that the development process of type 2 diabetes mellitus could be the following four stages according to the function of β cell: β cell function normal, β cell functional compensation, β cell function loss of compensation, and finally β cell function failure.

Research progress of treatments on colorectal cancer in elderly patients / 中华老年医学杂志

The patients aged ≥ 60 years account for 70％ of Chinese population with colorectal cancer.However,with the increase of age,the treatment methods such as surgery,radiotherapy,chemotherapy become inappropriate and impeding factors for the self-care of the elderly.Several clinical trials show that only a part of the elderly,who are in good health condition,can get the clinical benefits from these therapies.Therefore,this review focused on the comprehensive treatments for elderly patients with colorectal cancer,then reached the best individualized treatment,so as to improve the survival and quality of life.

Explore the occurrence and development of β cell dysfunction and insulin resistance according to the stratification on normal glucose tolerance / 中华内分泌代谢杂志

After the stratification of the normal glucose tolerance, the changes of insulin resistance and βcell function in the development of type 2 diabetes mellitus were investigated. A retrospective analysis on data of 275 cases with oral glucose insulin releasing tests. The area under the insulin curve (AUCINS ) 108. 43 mU/ L was taken as the critical value of diagnosis. Normal glucose tolerance subjects were divided into the NGT-a group(AUCINS<108. 43 mU/ L) and the NGT-b group(AUCINS≥108. 43 mU/ L). The plasma glucose, insulin, insulin sensitivity, and β cell function were compared among the 4 groups: NGT-a group (n=96), NGT-b group (n=49), prediabetes group (n=71), and type 2 diabetes mellitus group ( n = 59). Among the fasting insulin, 2 h insulin, AUCINS , early-phase insulin secretion index(△I30 / △G30), the ratio of total insulin area under curve, and total glucose area under curve, disposition index, homeostasis model assessment for insulin resistance, and Matsuda insulin sensitivity index, the relationship as follows: NGT-b group>prediabetes group>NGT-a group>type 2 diabetes mellitus group. The NGT-b group was always the highest, prediabetes group was lower, NGT-a group and type 2 diabetes mellitus group were the lowest, there were significant differences (all P<0. 05). Making the NGT-a group as the basic state, in the NGT-b group, β cell function has begun to appear compensation and insulin resistance, and β cell function compensation reached the peak, the β cell function in the prediabetes group was beginning to compensate for the deficiency, the function of β cell in type 2 diabetes mellitus group decreased further. These findings suggest that the development process of type 2 diabetes mellitus could be the following four stages according to the function of β cell: β cell function normal, β cell functional compensation, β cell function loss of compensation, and finally β cell function failure.

Role of epileptogenicity index in the epileptogenic zone and network localization / 中华神经科杂志

Objective To propose a novel stereo-electroencephalography(SEEG) quantitative measure analyzing ictal high frequency (60-90 Hz) and calculating high frequency epileptogenicity index (HFEI) to localize epileptogenic zone and evaluate epileptogenic network. Methods The clinical presurgical evaluation and SEEG data of 15 patients who were performed SEEG electrodes implantation from April 2015 to March 2016 were analyzed retrospectively. Post-implantation head CT images and 3D MRI data were fused for accurately identifying and locating each electrode contact. Ictal SEEG quantitative measure HFEI was calculated and threshold was set. The epileptogenic network was divided into focal, regional, multiple regional and bilateral ones and the results were compared with the pathological results.Results The epileptogenic network was focal for four patients, regional for four patients, multiple regional for six patients and bilateral for one patient (7/15). In terms of the pathology,two cases with hippocampal sclerosis both showed regional network. In four cases with cerebral malacia, two cases showed multiple regional network and the other two cases showed focal network. In six cases with cortical malformation, three cases showed multiple regional network, the other three cases showed focal, regional and bilateral networks respectively. Conclusions We explored a novel SEEG quantitative measure based on the high frequency power analysis,which is objective and could localize epileptogenic zone and evaluate the epileptic network.