Objective To evaluate the effect of stellate ganglion block (SGB) on postoperative synaptic structure in hippocampal CA3 region in aged rats.Methods Seventy-two male Sprague-Dawley rats,aged 20-22 months,weighing 550-650 g,were randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),operation group (group O) and SGB + operation group (group SGB).Group SGB received right SGB with 0.25％ bupivacaine 0.15 ml.Groups O and SGB underwent 30 min of exploratory laparotomy starting from 15 min after the end of administration.Y-maze test was performed on 1 day after operation in 6 rats chosen from each group for assessment of cognitive function.The frequency of standard training and standard time were recorded.Six rats were chosen from each group on 1,3 and 7 days after operation and sacrificed and the hippocampal CA3 region was isolated for microscopic examination and for measurement of synaptic structure.Results Compared with group C,the standard time was significantly prolonged,and the frequency of standard training was increased in groups O and SGB,the width of synaptic cleft was increased,the thickness of post-synaptic density was decreased,the length of active zones was shortened,and the curvature of the synaptic interface was decreased on 1,3 and 7 days after operation in group O (P ＜ 0.05),and no significant changes were found in each synaptic structure parameter in group SGB (P ＞ 0.05).Compared with group O,the standard time was significantly shortened,the frequency of standard training was decreased,the width of synaptic cleft was decreased,the thickness of the post-synaptic density was increased,the length of active zones was prolonged,and the curvature of the synaptic interface was increased on 1,3 and 7 days after operation in group SGB (P ＜ 0.05).Conclusion The mechanism by which SGB improves the postoperative cognitive dysfunction in aged rats may be related to inhibition of changes of synaptic structure in hippocampal CA3 region.

Background and purpose:MicroRNA (miRNA) belongs to a class of 19 to 30 nucleotide-long, endogenous noncoding RNA expressed in eukaryotes and predominantly inhibits gene expression at the post-transcriptional level. The miRNAs play critical roles in cell proliferation and differentiation, apoptosis, metabolism, and immune regulation. This study aimed to detect the expression of miR-216a-5p in lung cancer tissues and lung cancer cell lines, and to discuss the effects of miR-216a-5p on the invasion ability of lung cancer cells and the mechanism.Methods:Quantitative real-time PCR (qRT-PCR) was used to detect the expression of miR-216a-5p in lung cancer tissues of 55 cases and 7 lung cancer cell lines. Three lung cancer cell lines of A549, 95D and H460 were transiently transfected by miR-216a-5p, and Transwell was used to detect the effects of miR-216a-5p on the invasion of lung cancer cell lines. The dual luciferase reporter plasmids containing the miR-216a-5p candidate target gene and the gene of matrix metalloproteinase 16 (MMP16) were predicted and constructed. qRT-PCR and Western blot were used to detect the changes in mRNA and protein levels of target geneMMP16 by miR-216a-5p. The interference of MMP16 by siRNA and up-regulation miR-216a-5p by transfection were compared on the invasion of lung cancer cells.Results:The miR-216a-5p expression levels were all signiifcantly reduced in 90.91% (50 of 55 patients) tumor tissues compared with corresponding adjacent normal lung tissues (P<0.05). The miR-216a-5p expression levels were only 7.00%-32.00%in 7 lung cancer cells compared with the control group (P<0.05). Up-regulation of the expression of miR-216a-5p inhibited the invasion of lung cancer cells; interference of MMP16 by siRNA, as well as up-regulating miR-216a-5p by transfection, inhibited the expression of MMP16 in lung cancer leading to inhibition of the invasion of lung cancer cells. Conclusion:miR-216a-5p can be a candidate marker in clinical diagnosis and it can inhibit the invasion of lung cancer cells by down-regulating the expression of MMP16.

Objective To investigate the effect of stellate ganglion block (SGB) on the postoperative cognitive function in aged rats.Methods Sixty-four male Sprague-Dawley rats,aged 18-20 months,weighing 500-700 g,were randomly divided into 4 groups (n =16 each):control group (group C),operation group (group O),normal saline + operation group (group NS) and SGB + operation group (group SGB).Group SGB received right SGB with 0.25％ bupivacaine 0.15 ml,while the equal volume of normal saline was given in group NS.Groups O,SGB and NS underwent 30 min of exploratory laparetomy starting from 15 min after the end of administration.Morris water-maze test was performed at days 1-6 after operation in 10 rats chosen from each group.The escape latency and frequency of crossing the original platform were recorded.Two rats were chosen from each group at 1,2 and 3 d after operation and sacrificed and the hippocampi were removed for microscopic examination.Results Compared with group C,the escape latency wag significantly prolonged on 1-5 days after operation in groups O and NS,the escape latency wag significantly prolonged on 1 and 2 days after operation in group SGB,and the frequency of crossing the original platform was decreased in groups O,NS and SGB (P ＜ 0.05).Compared with group NS,the escape latency wag significantly shortened,and the frequency of crossing the original platform was increased on 1-5 days after operation in group SGB (P ＜ 0.05).The number of hippocampal neurons was significantly larger at 2 and 3 d after operation in group SGB than in group NS.Conclusion SGB can improve the postoperative cognitive function in aged rats.

Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.

Objective To evaluate the effects of age factors on hypothermia-induced reduction of ischemia-reperfusion(I/R)injury in isolated rat hearts.Methods Pathogen-free healthy aged male Sprague-Dawley rats,aged 18-20 months,weighing 400-600 g,and young rats,aged 4-6 months,weighing 280-350 g,were used in the study.After the animals were anesthetized with intraperitoneal chloral hydrate and heparinized,their hearts were excised and perfused with K-H solution in a Langendorff apparatus.Twenty-four isolated hearts of aged rats were assigned into 2 groups(n=12 each)using a random number table:I/R group(group AI/R)and hypothermia group(group AH).Twenty-four isolated hearts of young rats were assigned into 2 groups(n=12 each)using a random number table:I/R group(group YI/R)and hypothermia group(group YH).Perfusion was suspended for 30 min followed by 120 min reperfusion to establish the model of I/R.The temperature was maintained at 37 ℃ during the whole process in AI/R and YI/R groups.The hearts were perfused with 34 ℃ K-H solution until 120 min of reperfusion starting from onset of reperfusion in AH and YH groups.At 30 min of equilibration(T0)and 15,30,60 and 120 min of reperfusion(T1-4),heart rate(HR),left ventricular developed pressure(LVDP),the maximum rate of increase in left ventricular pressure(+dp/dtmax),and the minimum rate of increase in left ventricular pressure(+dp/dtmin)were recorded.Six hearts from each group were randomly selected at T4,and myocardial specimens were obtained for determination of ATP,superoxide dismutase(SOD)and malondialdehyde(MDA)levels and myocardial infarct size(IS).Results Compared with group YI/R,HR was significantly decreased at T1-4,ATP and SOD levels were increased,and the MDA content and myocardial IS were decreased in group YH,and the HR,LVDP,+dp/dtmax and +dp/dtmin at T0 and ATP and SOD levels at T4 were significantly decreased,and the MDA content and myocardial IS were increased in group AI/R(P<0.05).Compared with group YH,HR,LVDP,+dp/dtmax and +dp/dtmin at T0 and ATP and SOD levels at T4 were significantly decreased,and the MDA content and myocardial IS were increased in group AH(P<0.05).Compared with group AH,the levels of ATP and SOD were significantly decreased,and the MDA content and myocardial IS were increased in group AI/R(P<0.05).Conclusion Age factors affect the efficacy of hypothermia in reducing I/R injury in isolated rat hearts,and hypothermia provides better cardioprotection for young rats than for aged rats.

Objective To investigate he intervention effect of knockdown of activating transcription factor 4(ATF-4)on fructose-induced lipid accumulation in liver cells. Methods HepG2 cells were divided into the control group(C),high fructose group(F),high-fructose+negative control group(F+NC)and high-fructose+ATF-4 siRNA group(F+ATF-4-). The mRNA level of gens of the upstream transcriptional factors and ERS markers was detected. The protein level of ACC,FAS and SCD-1 was also detected. Results Compared with group C,the mRNA expression of SREBP-1c,ChREBP,GRP78 and CHOP was increased(P < 0.01),while ATF-4 knock-down decreased the expression of the above genes(P < 0.01 ,respectively). Compared with group C ,the protein expression of ACC,FAS and SCD-1 was increased in group F(P<0.01,respectively). While ATF-4 knockdown decreased the protein expression of ACC ,FAS and SCD-1. Conclusions ATF-4 knockdown can improve the lipid steatosis induced by fructose through down-regulating lipid lipogenesis ,indicating ATF-4 possesses a regulatory effect on lipogenesis.

Objective:To make a new simple respirator and observe the oxygen therapy effect of the respirator on patients with severe and critical coronavirus disease 2019 (COVID-19).Methods:Based on the infectivity and hospital requirements of COVID-19, a new simple respirator was designed by the medical staff of the Department of Anesthesiology of the Second Affiliated Hospital of Nanchang University, which was applied on the 22 patients with severe and critical COVID-19 who needed oxygen therapy admitted to the Cancer Center of Tongji Medical College of Huazhong University of Science and Technology from February 15th to March 15th in 2020. The new simple respirator contained two National Utility Model Patents (a respirator: ZL 2015 2 0410623.6, a fluid switch and oxygen suction device: ZL 2017 2 0873509.6), which was mainly composed of anesthesia mask and filter, L-shaped connecting tube, soft breathing bladder, connecting tube and elastic fixing belt. When in use, the anesthesia mask was fixed to the patient's mouth and nose with elastic straps, the connecting tube was inserted into the oxygen meter interface, the oxygen flow was adjusted to 6-10 L/min, and the L-shaped connecting tube was opened immediately after the soft breathing bag was full. The carbon dioxide and excess oxygen in the body was discharged from exhaust port. The oxygen flow was lowered to 2-3 L/min, the patient's respiratory rate (RR) was observed through the soft breathing bag fluctuations, and the oxygen flow was adjusted at any time. The changes of pulse oxygen saturation (SpO 2), RR and heart rate (HR) before and after application of new simple respirator were observed, and the blood gas test results of part of the patients were collected. Results:Twenty-two patients with severe and critical COVID-19 had significantly higher SpO 2 at 10 minutes after application of the new simple ventilator than before application (0.994±0.007 vs. 0.952±0.017, P < 0.01), and RR was significantly lower than that before application (times/min: 27.59±3.63 vs. 29.64±3.81, P < 0.01); after 1 day of application, each index was further improved. All 13 patients who received blood gas analysis indicated no carbon dioxide accumulation. Conclusions:The new simple respirator can significantly improve the oxygen therapy effect of patients with severe and critical COVID-19. At the same time, 2019 novel coronavirus (2019-nCoV) can be filtered through the filter to reduce the formation of aerosol and protect the medical staff and patients.

Objective To evaluate the role of sirtuin 1 ( SIRT1)∕nuclear factor kappa B ( NF-κB) signaling pathway in oxygen-glucose deprivation and restoration ( OGD∕R) injury to hippocampal neurons of mice. Methods The HT22 hippocampal neurons were seeded in a culture plate ( 96-well plate, 100 μl∕well; 6-well plate, 2 ml∕well) at the density of 5×104 cells∕ml or in a culture dish (6 cm in diameter), and then divided into 4 groups ( n=24 each) using a random number table method: control group ( group C) , OGD∕R group ( group OGD) , SIRT1 inhibitor EX-527 preconditioning group ( group EX) and SIRT1 agonist SRT1720 preconditioning group ( group SRT) . Neurons were cultured in normal culture atmosphere at 37 ℃ in group C. In OGD, EX and SRT groups, the culture medium was replaced with oxygen-poor flu-id, and neurons were exposed to 5％ CO2-95％ N2 for 12 h in an incubator at 37℃, oxygen-poor fluid was replaced with the culture medium, and neurons were cultured for 24 h in normal culture atmosphere at 37℃. SIRT1 inhibitor EX-5271 μmol∕L and SIRT1 agonist SRT172010 μmol∕L were added at 12 h beforeOGD in EX and SRT groups, respectively. The cell viability was measured by CCK8 assay, the activity of LDH was detected by chemical colorimetry, cell apoptosis rate was determined by flow cytometry, the ex-pression of SIRT1, NF-κB, IκBα, Bcl-2 and Bax was detected by Western blot, and the Bcl-2∕Bax ratio was calculated. Results Compared with group C, the cell viability was significantly decreased, LDH ac-tivity and cell apoptosis rate were increased, the expression of SIRT1, IκBα and Bcl-2 was down-regula-ted, the expression of NF-κB and Bax was up-regulated, and the Bcl-2∕Bax ratio was decreased in OGD, EX and SRT groups ( P<0. 05) . Compared with group OGD, the cell viability was significantly increased, the LDH activity and cell apoptosis rate were decreased, the expression of SIRT1, IκBαand Bcl-2 was up-regulated, the expression of NF-κB and Bax was down-regulated, and the Bcl-2∕Bax ratio was increased in group SRT, and the cell viability was significantly decreased, LDH activity and cell apoptosis rate were in-creased, the expression of SIRT1, IκBαand Bcl-2 was down-regulated, the expression of NF-κB and Bax was up-regulated, and the Bcl-2∕Bax ratio was decreased in group EX (P<0. 05). Conclusion SIRT1∕NF-κB signaling pathway inhibition is involved in OGD∕R injury to hippocampal neurons of mice.

Objective To analyze the role of NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome in a therapeutic mild hypothermia(34℃)treated isolated rat myocardial ischemia-reperfusion model and explore its mechanism.Methods Sixty clean grade adult male SD rats,aged 7-10 weeks,weighing 250-300 g.Using a random number table method,the rats were divid-ed into five groups:blank control group(group S),myocardial ischemia-reperfusion group(group IR),34℃mild hypothermia post-treated myocardial ischemia-reperfusion group(group MH),34℃mild hypother-mia post-treated myocardial ischemia-reperfusion+3-TYP group(group HT),and 34℃mild hypothermia post-treated myocardial ischemia-reperfusion+3-TYP+MCC950 group(group HTM),12 rats in each group.Group S perfused the rat heart at 37℃with a balanced perfusion solution for 180 minutes.Group IR re-ceived balanced perfusion of the rat heart at 37℃for 30 minutes,followed by ischemia for 30 minutes and reperfusion with 37℃perfusion for 120 minutes.Group MH perfused the rat heart at 37℃for 30 minutes,followed by ischemia for 30 minutes and reperfusion with 34℃perfusion solution for 120 minutes.Group HT perfused the hearts of rats at 37℃for 30 minutes,followed by ischemia for 30 minutes,silent mating type information regulation 2 homolog 3(sirt3)inhibitor 3-TYP was added to the perfusate,and then per-fused at 34℃for 120 minutes.Group HTM perfused the hearts of rats at 37℃for 30 minutes,followed by ischemia for 30 minutes,sirt3 inhibitor 3-TYP and NLRP3 inhibitor MCC950 were added to the perfusate,and then perfused at 34℃for 120 minutes.The isolated heart was obtained 120 minutes after reperfusion,and the concentrations of IL-6 and IL-1β in the perfused cardiac fluid was measured using ELISA method,Western blot method for detecting the relative content of NLRP3 and sirt3 proteins in myocardial tissue,1%triphenyl tetrazolium chloride staining for calculating myocardial infarction area,and HE staining for observ-ing myocardial pathological changes.Results Compared with group S,HR were significantly slowed down,LVSP,±dp/dtmax were significantly decreased,and LVEDP were significantly increased 30,60,90,and 120 minutes after reperfusion,the concentrations of IL-6 and IL-1β in cardiac fluid leakage,and the per-centage of myocardial infarction area were significantly increased in groups IR,MH,HT,and HTM(P＜0.05),the content of sirt3 protein in myocardial tissue were significantly reduced,while the content of NLRP3 protein were significantly increased in groups IR,HT,and HTM(P＜0.05),the contents of sirt3 and NLRP3 protein in the myocardial tissue were significantly increased in group MH(P＜0.05).Com-pared with group IR,HR were significantly increased,LVSP,±dp/dtmax were significantly increased,and LVEDP were significantly decreased 30,60,90,and 120 minutes after reperfusion,the concentrations of IL-6 and IL-1β in cardiac fluid leakage and the percentage of myocardial infarction area were significantly decreased in groups MH and HTM(P＜0.05),the content of sirt3 protein in myocardial tissue was signifi-cantly increased,while the content of NLRP3 protein was significantly decreased in group MH(P＜0.05),the content of NLRP3 protein in myocardial tissue was significantly reduced in group HTM(P＜0.05).Compared with group MH,HR were significantly slowed down,LVSP,±dp/dtmax were significantly de-creased,and LVEDP were significantly increased 30,60,90,and 120 minutes after reperfusion,the con-centrations of IL-6 and IL-1β in cardiac fluid leakage,the percentage of myocardial infarction area,and the content of NLRP3 protein in myocardial tissue were significantly increased in group HT(P＜0.05),the content of sirt3 protein in myocardial tissue was significantly reduced in groups HT and HTM(P＜0.05).Compared with group HT,HR were significantly increased,LVSP,±dp/dtmax were significantly increased,and LVEDP were significantly decreased 30,60,90,and 120 minutes after reperfusion,the concentrations of IL-6 and IL-1β in cardiac fluid leakage,the percentage of myocardial infarction area,and the content of NLRP3 protein in myocardial tissue were significantly reduced in group HTM(P＜0.05).Conclusion Therapeutic mild hypothermia(34℃)can improve hemodynamic parameters of isolated hearts and reduce the concentrations of IL-6 and IL-1β,NLRP3 protein content in myocardial tissue,percentage of myocardial infarction area,improve myocardial pathological changes,and reduce myocardial ischemia-reperfusion injury in rats,the mechanism may be related to the mitochondrial mediated sirt3 pathway inhibiting the high expres-sion of inflammatory corpuscle NLRP3.

Objective: To describe the MICM (morphology, immunology, cytogenetics and molecular biology) characteristics of a case of acute myelomonocytic leukemia M 4C . Methods: The medical history data of the case of M 4C admitted to our hospital was reviewed. The results of bone marrow cell morphology, cytochemical stains, bone marrow biopsy, immunophenotype, cytogenetics, molecular test and NGS (next-generation sequencing) of the case were analyzed. Results: The bone marrow smear showed markedly active proliferation of bone marrow cells in which the myelomonocytic cells accounted for 85.6%. Cytochemical stains showed peroxidase (POX) stain partially and weakly positive; specific esterase AS-DCE partially positive; non-specific esterase α-NBE partially positive and smothered by sodium fluoride; non-specific esterase AS-DAE partially positive and smothered by sodium fluoride. Bone marrow biopsy showed hyperproliferative cells and diffused hyperplasia of blasts. Immunophenotype analysis showed that the abnormal cell population was positive for CD11B, CD64, CD56, cMPO, CD33, CD41, CD61, CD38 and CD58, but negative for CD13, CD34, CD117, CD7, CD123, HLA-DR, CD10, CD19, CD20, CD2, CD14, CD235, CD15, CD303, CD304, CD25, cCD79a, cCD3, cCD22, CD1a and TDT. Cytogenetic analysis showed 47, XY, t(9;11) (p22;q23),+mar. The molecular test for leukemia showed MLLT3/KMT2A gene rearrangement. NGS showed NRAS and TET2 mutation. The case was finally diagnosed as AML (acute myelomonocytic leukemia) M 4C with t(9;11)(p22;q23), MLLT3-KMT2A. Conclusion Leukemia M 4C may show the characteristics of both granulocytes and monocytes with complex morphological features. The combined examination of MICM should be necessary for the diagnosis of M 4C with great significance.