Objective To investigate the clinical and pathologic features of patients with gastric neuroendocrine neoplasms (g-NENs).Methods A total of 50 cases of g-NENs diagnosed by pathology in the First Hospital of Jilin University from May 2012 to January 2016 were retrospective analyzed to summarize the clinical manifestations and pathological features.The location of lesion,gross morphology,maximum diameter,lymph node metastasis rate,distant metastasis rate,and survival time between patients with neuroendocrine tumors and neuroendocrine carcinomas were compared.Results Among the 50 patients,34 were males and 16 were females with a male to female ratio of 2.125 ∶ 1.Their age ranged from 33 to 77 years with an mean age of 60± 11 years.There were 13 cases (26％) of neuroendocrine tumors,31 (62％) neuroendocrine carcinomas,and 6 (12％) mixed adenoneuroendocrine carcinomas.The maximum diameter of lesion was less than 2 cm in 8 cases (16％),and equal or greater than 2 cm in 42 cases (84％).There was single lesion in 48 cases (96％) and multiple lesions in 2 cases (4％).There were 19 cases (38％) located in gastric antrum,16 (32％) in gastric body,11 (22％) in gastric fundus or cardia,3 (6％) in gastric angle,and 1 (2％) in the junction of gastric antrum and body.Thirty-two patients (64％) had metastasis,including 20 cases of lymph node metastasis and 12 cases of distant metastasis.The clinical symptoms of the patients were different,mainly were digestive system symptoms and tumor occupying symptoms,and no carcinoid syndrome was reported.The gross morphology (x2 =24.446.P =0.000).maximum diameter (t =-4.044,P =0.001),lymph node metastasis rate (x2=4.435,P =0.035),and survival time (t =2.925,P =0.000) were significantly differences between 13 cases of neuroendocrine tumors and 37 cases of neuroendocrine carcinoma.But the location of lesion (x2 =6.921,P=0.082) and distant metastasis rate (x2 =0.715,P =0.389) were no statistically different between the two groups.Conclusion Gastric neuroendocrine neoplasms can occur in any part of stomach,majority of tumor is single lesion and lack of specific clinical manifestations.The mostly gross morphology of gastric neuroendocrine carcinoma and mixed adenoneuro-endocrine carcinoma are ulcer type with a large diameter and poor prognosis.

Objective To evaluate the effect of cardamonin on acute lung injury induced by hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty-two male Sprague-Dawley rats,aged 18-24 weeks,weighing 200-250 g,were divided into 4 groups (n =8 each) using a random number table:sham operation group (group Sham);group HSR;cardamonin group (group CA);cardamonin + adenosina A2A receptor antagonist ZM241385 group (group CZM).Bilateral common carotid arteries were only cannulated in group Sham.The left common carotid artery was cannulated for blood-letting until mean arterial pressure was reduced to 35-45 mmHg and maintained at this level for 30 min,and the animals were then resuscitated by infusion of shed blood and normal saline two-fold volume of shed blood to establish HSR model in HSR,CA and CZM groups.ZM241385 5 mg/kg was injected intraperitoneally at 30 min before blood-letting in group CZM,and cardamonin 75 mg/kg was injected intraperitoneally immediately after the beginning of resuscitation in CA and CZM groups.The rats were sacrificed at 2 h after completion of resuscitation,bronchoalveolar lavage fluid (BALF) was collected for determination of neutrophil count,and lungs were removed for microscopic examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),contents of tumor necrosis factor-alpha (TNF-ct),interleukin-1 (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay) and expression of adenosine A2A receptors in lung tissues (by Western blot).Results Compared with group Sham,the neutrophil count in BALF,W/D ratio and contents of TNF-α,IL-1β and IL-6 were significantly increased,the expression of adenosine A2A receptors was significantly down-regulated in group HSR,and the neutrophil count in BALF and contents of TNF-α and IL-6 were significantly increased (P＜0.05),and no significant changes were found in W/D ratio,content of IL-1β,and expression of adenosine A2A receptors in group CA (P＞0.05).Compared with group HSR,the neutrophil count in BALF,W/D ratio and contents of TNF-α,IL-1β and IL-6 were significantly decreased,the expression of adenosine A2A receptors was significantly up-regulated (P＜0.05),and the pathological changes were significantly attenuated in group CA,and no significant changes were found in the parameters mentioned above in group CZM (P＞0.05).Compared with group CA,the neutrophil count in BALF,W/D ratio and contents of TNF-α,IL-1β and IL-6 were significantly increased,the expression of adenosine A2A receptors was significantly down-regulated (P＜0.05),and the pathological changes were aggravated in group CZM.Conclusion Cardamonin can attenuate acute lung injury induced by HSR in rats,and activated adenosine A2A receptors and inhibited inflammatory responses are involved in the mechanism.

Objective To establish a quantitative method for the simultaneous measurement of the residual level of three pesticides in Nelumbinis semen by GC-MS/MS. Methods The samples were extracted by acetonitrile and purify by Cleanert TPH column. The samples were then tested by GC-MS/MS. Information on relative retention time and mass charge ratio was used for qualitative analysis. The peak area obtained by secondary ion MS of bifenthrin (181.1/166.1)was used as the reference peak to calculate the relative correction factor for the peak area of fenpropathrin (265.1/210.1) and deltamethrin (252.9/93.0), to establish a method using bifenthrin as the reference substance to determinate there sidual quantity of three pesticides in Nelumbinis semen by GC-MS/MS. Results When the injection quantity of the sample containing bifenthrin,fenpropathrin and deltamethrin in the range of 0.01-0.1 ng , there was a good linear relationship between the injection quantity and peak are a Limitation of quantification (LOQ) of bifenthrin , fenpropathrin and deltamethrin were 4.321×10-4 ng, 3.435×10-4 ng, 8.913×10-3 ng, respectively. The average recovery rates of bifenthrin, fenpropathrin and deltamethrin were 93.5％, 93.5％ and 93.8％, respectively. Conclusions The method of quantitative analysis of multi-components with a single-marker is simple, quick and accurate. It suitable for the detection of residual quantity of bifenthrin, fenpropathrin and deltamethrin in Nelumbinis semen.

Objective To investigate the effect of anesthesia factor on lung injury in patients un?dergoing thoracoscopic radical lung cancer surgery and to evaluate efficacy of combination of thoracic para?vertebral block(TPVB)with dexmedetomidine mixed with ropivacaine and general anesthesia. Methods One hundred patients of both sexes, aged 18-64 yr, with body mass index of 18-25 kg∕m2, of American Society of Anesthesiologists physical statusⅡorⅢ, scheduled for elective thoracoscopic radical lung cancer surgery, were divided into 5 groups(n=20 each)using a random number table: general anesthesia group (group G), TPVB with ropivacaine combined with general anesthesia group(group R), intravenously in?fused dexmedetomidine combined with general anesthesia group(group Div), intravenously infused dexme?detomidine plus TPVB with ropivacaine combined with general anesthesia group(group Div+R), and TPVB with dexmedetomidine mixed with ropivacaine combined with general anesthesia group(group Dtp+R). In group R, TPVB was performed under ultrasound guidance, two?point method was selected accord?ing to the position of intercostal space at surgical incision, and 0.5% ropivacaine 10 ml was injected into each puncture site. Dexmedetomidine 0.5 μg∕kg was intravenously infused over 10 min in group Div. Dexmedetomidine was intravenously infused for TPVB in group Div+R. TPVB solution contained dexmedeto?midine 0.5 μg∕kg and ropivacaine in group Dtp+R. Anesthesia was then induced and maintained by IV in?fusion of propofol and remifentanil. The intraoperative consumption of propofol and remifentanil and develop?ment of adverse reactions such as hypoxemia, hypotension and bradycardia were recorded. Normal lung tis?sues around the tumor margin were obtained immediately after tumor resection for determination of the ex?pression of hypoxia?inducible factor 1 alpha(HIF?1α), BCL2∕adenovirus E1B 19kDa interacting protein 3 (BNIP3)and microtubule?associated protein 1 light chain 3Ⅱ(LC3Ⅱ)(by Western blot), contents of tumor necrosis factor?alpha(TNF?α)and interleukin?6(IL?6)in lung tissues(by enzyme?linked immu?nosorbent assay)and cell apoptosis(by TUNEL)and for examination of the pathological changes(with a light microscope)which were scored. Apoptosis index was calculated. Results The amount of propofol consumed was significantly lower in Div+R and Dtp+R groups than in the other three groups, and the a?mount of remifentanil consumed was significantly higher in G and Div groups than in the other three groups (P<0.05). The incidence of hypertension and tachycardia was significantly lower in R and Div groups than in group G(P<0.05). The incidence of hypotension was significantly lower in R, Div and Dtp+R groups than in group Div+R(P<0.05). The incidence of bradycardia was significantly higher in Div and Div+R groups than in group R(P<0.05). Compared with G and R groups, apoptosis index, contents of TNF?α and IL?6 and lung injury scores were significantly decreased, and the expression of HIF?1α, BNIP3 and LC3Ⅱ was up?regulated in Div, Div+R and Dtp+R groups(P<0.05). Compared with group Div, the TNF?α content and lung injury scores were significantly decreased, and the expression of HIF?1α and LC3Ⅱwas up?regulated in Div+R and Dtp+R groups, and the IL?6 content was significantly decreased in group Dtp+R(P<0.05). Conclusion Combination of TPVB with dexmedetomidine mixed with ropivacaine and general anesthesia produces better efficacy in reducing lung injury in patients undergoing thoracoscopic radi?cal lung cancer surgery.

Objective To evaluate the efficacy of different doses of dexmedetomidine mixed with ropivacaine used for thoracic paravertebral nerve block (TPVB) combined with general anesthesia in the pa-tients undergoing radical resection of lung cancer. Methods One hundred patients of both sexes, aged 18-64 yr, with body mass index of 18-25 kg∕m2, of American Society of Anesthesiologists physical statusⅡorⅢ, scheduled for elective radical resection of lung cancer, were divided into 5 groups (n＝20 each) using a random number table method: general anesthesia group ( group G), TPVB with ropivacaine com-bined with general anesthesia group (group R), TPVB with 0. 5 μg∕kg dexmedetomidine mixed with ropiva- caine combined with general anesthesia group (group RD0. 5), TPVB with 1. 0 μg∕kg dexmedetomidine mixed with ropivacaine combined with general anesthesia group (group RD1. 0), and TPVB with 2. 0 μg∕kg dexmedetomidine mixed with ropivacaine combined with general anesthesia group ( group RD2. 0). Two-point TPVB was performed, and anesthetic 10 ml was injected into each puncture site. Zero point five per-cent ropivacaine was administered in group R. Dexmedetomidine 0. 5, 1. 0 and 2. 0 μg∕kg mixed with ropi-vacaine at the final concentration of 0. 5% were injected in group RD0. 5, group RD1. 0 and group RD2. 0, respectively. Anesthesia was maintained by intravenously infusing propofol and remifentanil and cisatracuri-um was intermittently injected to maintain muscle relaxation. The occurrence of intraoperative hypotension and bradycardia was recorded. The emergence time, extubation time, duration of postanesthesia care unit (PACU) stay and requirement for rescue analgesia during emergence from anesthesia were also recorded. Small marginal lung samples next to the tumor were harvested immediately after removing the tumor tissues. The expression of caspase-3 and Bcl-2 was determined by Western blot, the content of IL-1β in lung tissues was determined by enzyme-linked immunosorbent assay, the cell apoptosis was evaluated by TUNEL, and apoptosis index ( AI) was calculated. The injury to lung tissues was observed with a light microscope and scored. Results Compared with G and R groups, AI, IL-1β content and lung injury score were signifi-cantly decreased, the expression of caspase-3 was down-regulated, and the expression of Bcl-2 was up-regulated in the other three groups, the incidence of hypotension and bradycardia was significantly in-creased in group RD1. 0, the incidence of bradycardia was significantly decreased in group RD0. 5, and the incidence of hypotension and bradycardia was significantly increased, and the emergence time, extu-bation time and duration of PACU stay were prolonged in group RD2. 0 ( P＜0. 05). Compared with group RD0. 5, IL-1β content was significantly decreased, and the incidence of hypotension and bradycardia was increased in group RD1. 0, and IL-1β content was significantly decreased, the incidence of hypoten-sion and bradycardia was increased, and the emergence time, extubation time and duration of PACU stay were prolonged in group RD2. 0 ( P＜0. 05). Compared with group RD1. 0, the incidence of hypotension and bradycardia was significantly increased, and the emergence time, extubation time and duration of PACU stay were prolonged in group RD2. 0 ( P＜0. 05). There was no significant difference in the AI, expression of caspase-3 and Bcl-2, lung injury score, or requirement for rescue analgesia among RD0. 5, RD1. 0 and RD2. 0 groups ( P＞0. 05). Conclusion Dexmedetomidine 0. 5 μg∕kg mixed with ropiva-caine for TPVB combined with general anesthesia provides better efficacy in the patients undergoing radical resection of lung cancer.

Objective:To investigate effect of reperfusion injury following different ischemic duration on skeletal muscle in rats.Methods:A model of ischemia/reperfusion injury (IRI) was established by unilateral clamping femoral artery and additional application of tourniquet in skeletal muscle of hind limbs in 35 male Wsitar rats. According to different ischemia time, the animals were assigned to 2-hour ischemia and 24-hour reperfusion (I2R24 group), 2.5-hour ischemia and 24-hour reperfusion (I2.5R24 group), 3-hour ischemia and 24-hour reperfusion (I3R24 group), 4-hour ischemia and 24-hour reperfusion (I4R24 group) and sham group, with 7 rats per group. At the end of reperfusion, gastrocnemious tissues and plasma samples were collected and analyzed. The ratio of wet ∶ dry weight (W/D) was used to measure muscle edema. The assay of 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was conducted to evaluate muscle viability. HE staining was executed to observe histopathological changes. Immunofluorescence staining was performed to assess the levels of C1q, C3b/c, tissue factor (TF), fibrinogen (FN), bradykinin receptor 1 (BR1), BR2, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, fibrinogen-like protein-2 (FGL-2) and myeloperoxidase (MPO) in muscle tissues. ELISA method was used to determine the concentrations of interferonγ (IFN-γ), interleukin7 (IL-7), IL-18, macrophage inflammatory1α (MIP-1α) and monocyte chemotactic protein 1 (MCP-1) in plasma.Results:With prolongation of ischemia time and subsequent reperfusion, tissue edema became severe gradually. The ratio of W/D was 5.3±0.2, 6.1±0.3, 6.9±0.2, 7.6±0.3 in I2R24, I2.5R24, I3R24 and I4R24 groups, higher than that in sham group (4.5±0.1) (all P<0.01). Muscle viability got decreased gradually. Muscle viability was (62.4±3.5)%, (45.3±3.3)%, (35.4±3.4)%, (27.1±5.9)% in I2R24, I2.5R24, I3R24 and I4R24 groups, lower than that in sham group［(93.8±7.2)%］(all P<0.01). Histopathological changes became aggravated gradually. The most severe group was I4R24 group, with the most severe myocyte injury, interstitial edema and extensive inflammatory infiltration, followed by I3R24, I2.5R24 and I2R24 groups in order. There was normal structure integrity and neatly arranged myocyte in sham group. Meanwhile, levels of C1q, C3b, FN, BR1, VCAM-1, E-selectin and FGL-2 got increased gradually. The highest levels for these factors were seen in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. The rough ratio of the number of positive MPO cells/total cell number under high lens (×200) were increased gradually, with the highest level in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. However, expression of TF and BR2 were not altered significantly among the groups. Plasma levels of INF-γ, IL-7, IL-18, MIP-1α and MCP-1 elevated gradually with prolongation of ischemia time (all P<0.01). The sequence was the sham group, I2R24 group, I2.5R24 group, I3R24 group and I4R24 groups for levels of these factors from low to high (all P<0.01). Conclusion:Reperfusion after prolongation of ischemia duration can increase the activation of complement, coagulation, kinin and endothelial cells as well as the release of inflammatory factors, and thus aggravate the degree of skeletal muscle tissue injury.

Objective To investigate the effects of air pollution on non-accidental death of residents in Yancheng City. Methods Data of daily air pollutions (PM_2.5, PM₁₀, NO₂, SO₂ and O₃), average temperature, average relative humidity, and daily death information during 2014-2019 were collected. The time series semi parametric generalized additive model was used to analyze the impact of short-term exposure of air pollutants on non-accidental death in Yancheng City. Results The average daily non-accidental deaths of the entire population, respiratory system, and circulatory system were 154, 25, and 51, respectively. With the increase of 10 µg/m³ SO₂, the risk of the estimated non-accidental death and respiratory death was increased by 1.19% (95%CI: 0.26%-2.12%) and 2.37% (95%CI: 0.65%-4.12 %), respectively. With the increase of 10 µ g/m³ NO₂, the risk of the estimated non-accidental death, circulatory system death and respiratory death was increased by 1.50%（95%CI:0.94%-2.05%）, 1.11%（95%CI:0.08%-2.16%）, and 1.53%（95%CI:0.71%-2.36%）, respectively. With the increase of 10 µg/m³ O₃, the risk of the estimated non-accidental death, circulatory system death and respiratory death was increased by 0.64%（95%CI: 0.25%-1.04%）, 0.81%（95%CI: 0.04%-1.58%）, and 0.78%（95%CI: 0.18%-1.37%）, respectively. Conclusion The short-term exposure of air pollutants affects the non-accidental death of the residents in Yancheng, and there are lag effects, of which NO₂, SO₂ and O₃ have a greater impact.

Objective:To compare the curative effects of different venous cannulas and drainage to improve patient's whole body oxygenation during the auxiliary process of venous-arterial extracorporeal membrane oxygenation (VA-ECMO) in lung transplantation.Methods:From December 2016 to December 2019, 12 patients who were assisted by VA-ECMO in one lung transplantation in People's Hospital of Henan Province were selected as the research objects. According to the number of side holes of venous cannulas, they were divided into two groups: one group with few side holes and other group with multiple side holes. The differences in blood gas indexes among the right radial artery, left radial artery, and right internal jugular vein before and after assistance were compared, and the assistance effect was evaluated.Results:The arterial partial pressure of oxygen (PaO 2) of blood gas indexes of the right and left radial arteries in both groups were significantly higher than that before assistance [mmHg (1 mmHg = 0.133 kPa): right and left radial artery in few side holes group: 79.5±4.2 vs. 48.3±3.8 and 88.1±3.5 vs. 48.3±3.8; right and left radial artery in multiple side holes group: 67.7±5.9 vs. 48.7±3.2 and 84.0±3.8 vs. 48.7±3.2, all P < 0.05]. The arterial partial pressure of carbon dioxide (PaCO 2) of blood gas index was significantly lower than that before assistance (mmHg: 44.2±2.6 vs. 71.7±4.4 for the right radial artery and 44.7±1.4 vs. 71.7±4.4 for the left radial artery in the group with few side holes; 46.2±2.1 vs. 71.2±3.5 for the right radial artery and 44.1±1.9 vs. 71.2±3.5 for the left radial artery in the group with multiple side holes, all P < 0.05). The partial pressure of oxygen in venous blood (PvO 2) of blood gas index of ECMO system in the group with few side holes was significantly lower than that of the multiport side holes group (mmHg: 56.4±3.2 vs. 88.7±1.5, P < 0.01), and the partial pressure of carbon dioxide in venous blood (PvCO 2) was significantly higher than that of multiport side holes group (mmHg: 63.6±3.7 vs. 44.2±1.7, P < 0.01). Conclusions:When VA-ECMO is used in lung transplantation, the superior vena cava blood flow can be fully drained by using intravenous cannula with few side holes. It can effectively improve the oxygenation of the upper body of lung transplant patients, avoid the dilemma of hypoxemia in the upper body and hyperxemia in the lower body, provide more effective assistance to patients undergoing single lung transplantation, and is more meaningful for improving the oxygenation status of the whole body in patients undergoing single lung transplantation.

Objective:To investigate the influence of long non-coding RNA (lncRNA) DIO3OS in ketamine-induced neurotoxicity and its mechanism in mouse hippocampal neurons.Methods:(1) Primary mouse hippocampal neurons were isolated and cultured; CCK-8 assay was used to detect the viability of cells treated with different concentrations of ketamine (0, 25, 50, 100, 200 μmol/L), and qPCR was used to detect DIO3OS mRNA expression. (2) Hippocampal neurons were divided into 4 groups: control group, ketamine group (cultured with 50 μmol/L ketamine for 24 h), ketamine+pc group (transfected with pcDNA3.1 plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h), and ketamine+DIO3OS group (transfected with pcDNA3.1-DIO3OS plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h); cell viability was detected by CCK-8 assay; lactic dehydrogenase (LDH) release was determined by LDH cytotoxicity assay kit; cell apoptosis was detected by flow cytometry; mRNA expressions of DIO3OS and brain-derived neurotrophic factor ( BDNF) were examined by qPCR; protein expressions of polypyrimidine tract-binding protein 1 (PTBP1) and BDNF were detected by Western blotting. (3) Total proteins of routinely cultured neurons were extracted, and RNA Pull-Down assay was used to detect whether DIO3OS mRNA and BDNF mRNA could directly bind to PTBP1 protein. (4) Hippocampal neurons were divided into ketamine+DIO3OS+si-NC group (co-transfected with pcDNA3.1-DIO3OS plasmid and si-NC plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h) and ketamine+DIO3OS+si-PTBP1 group (co-transfected with pcDNA3.1-DIO3OS plasmid and si-PTBP1 plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h); qPCR was used to examine the mRNA expressions of DIO3OS and BDNF; Western blotting was used to detect the protein levels of PTBP1 and BDNF. (5) Hippocampal neurons were divided into ketamine group (cultured with 50 μmol/L ketamine for 24 h), ketamine+si-DIO3OS group (transfected with si-DIO3OS plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h), ketamine+si-PTBP1 group (transfected with si-PTBP1 plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h), and ketamine+si-DIO3OS+si-PTBP1 group (co-transfected with si-DIO3OS plasmid and si-PTBP1 plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h); qPCR was used to examine the mRNA expressions of DIO3OS and BDNF; Western blotting was used to detect the BDNF protein expression. (6) Hippocampal neurons were divided into ketamine+DIO3OS+si-NC group (co-transfected with pcDNA3.1-DIO3OS plasmid and si-NC plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h), ketamine+DIO3OS+si-BDNF group (co-transfected with pcDNA3.1-DIO3OS plasmid and si-BDNF plasmid for 48 h, and then cultured with 50 μmol/L ketamine for 24 h); cell viability was detected by CCK-8 assay; LDH release was determined by LDH cytotoxicity assay kit; cell apoptosis was detected by flow cytometry. Results:(1) Compared with 0 μmol/L ketamine, 25, 50, 100 and 200 μmol/L ketamine could significantly inhibit the cell viability and DIO3OS mRNA expression ( P<0.05). (2) Compared with control group, ketamine group had significantly decreased DIO3OS mRNA expression and cell viability, significantly increased LDH release and apoptotic rate, and statistically inhibited BDNF mRNA and protein expressions and PTBP1 protein expression ( P<0.05); compared with ketamine+pc group, ketamine+DIO3OS group had significantly increased DIO3OS mRNA expression and cell viability, significantly decreased LDH release and apoptotic rate, significantly elevated BDNF mRNA and protein expressions ( P<0.05). (3) RNA Pull-Down assay showed that Bio-labeled DIO3OS (Bio-DIO3OS) and Bio-labeled BDNF (Bio-BDNF) could adsorb PTBP1 protein, while Bio-labeled antisense strands of DIO3OS or BDNF (Bio-DIO3OS-AS and Bio-BDNF-AS) could not adsorb PTBP1 protein. (4) Compared with ketamine+DIO3OS+si-NC group, ketamine+DIO3OS+si-PTBP1 group had significantly inhibited BDNF mRNA and protein expressions and PTBP1 protein expression ( P<0.05); no significant difference was noted in DIO3OS mRNA expression ( P>0.05). (5) Compared with ketamine group, ketamine+si-DIO3OS and ketamine+si-DIO3OS+si-PTBP1 groups had significantly decreased DIO3OS mRNA expression ( P<0.05); compared with ketamine group, ketamine+si-DIO3OS and ketamine+si-PTBP1 groups had significantly decreased BDNF mRNA and protein expressions ( P<0.05); compared with ketamine+si-DIO3OS and ketamine+si-PTBP1 groups, ketamine+si-DIO3OS+si-PTBP1 group had significantly elevated BDNF mRNA and protein expressions ( P<0.05). (6) Compared with ketamine+DIO3OS+si-NC group, ketamine+DIO3OS+si-BDNF group had significantly reduced cell viability, and significantly increased LDH release and apoptotic rate ( P<0.05). Conclusion:LncRNA DIO3OS expression is decreased in ketamine-induced primary mouse hippocampal neurons; DIO3OS overexpression can alleviate ketamine-induced neurotoxicity in mouse hippocampal neurons by regulating BDNF expression via binding to PTBP1.

Mitophagy is an essential intracellular process that eliminates dysfunctional mitochondria and maintains cellular homeostasis. Mitophagy is regulated by the post-translational modification of mitophagy receptors. Fun14 domain-containing protein 1 (FUNDC1) was reported to be a new receptor for hypoxia-induced mitophagy in mammalian cells and interact with microtubule-associated protein light chain 3 beta (LC3B) through its LC3 interaction region (LIR). Moreover, the phosphorylation modification of FUNDC1 affects its binding affinity for LC3B and regulates selective mitophagy. However, the structural basis of this regulation mechanism remains unclear. Here, we present the crystal structure of LC3B in complex with a FUNDC1 LIR peptide phosphorylated at Ser17 (pS), demonstrating the key residues of LC3B for the specific recognition of the phosphorylated or dephosphorylated FUNDC1. Intriguingly, the side chain of LC3B Lys49 shifts remarkably and forms a hydrogen bond and electrostatic interaction with the phosphate group of FUNDC1 pS. Alternatively, phosphorylated Tyr18 (pY) and Ser13 (pS) in FUNDC1 significantly obstruct their interaction with the hydrophobic pocket and Arg10 of LC3B, respectively. Structural observations are further validated by mutation and isothermal titration calorimetry (ITC) assays. Therefore, our structural and biochemical results reveal a working model for the specific recognition of FUNDC1 by LC3B and imply that the reversible phosphorylation modification of mitophagy receptors may be a switch for selective mitophagy.
Crystallography, X-Ray ; Membrane Proteins ; chemistry ; metabolism ; Microtubule-Associated Proteins ; chemistry ; metabolism ; Mitochondrial Degradation ; Mitochondrial Proteins ; chemistry ; metabolism ; Peptides ; chemistry ; metabolism ; Phosphorylation ; Protein Structure, Quaternary