Objective To study the effect of nicorandil on the function of coronary artery endothelium during hypoxiareoxygenation.Methods Forty-five fresh porcine left anterior descending coronary artery rings in 2mm long were randomly divided into five groups.Control group ( n =9):incubation in Krebs-Henseleit (KH) at 37℃ for 90 minutes with a constant supply of oxygen ; Group A ( n =9):30-minute hypoxia ( PO2 ＜ 15 mm Hg) followed by 30 minutes reoxygenation in KH at 37℃ ; Group B ( n =9):60-minute hypoxia followed by 30 minutes reoxygenation in KH at 37 ℃ ; Group C ( n =9):60-minute hypoxia followed by 30 minutes reoxygenation in KH added nicorandil ( 0.2 μmol/L) at 37 ℃ ; Group D ( n =9 ):60-minute hypoxia followed by 30 minutes reoxygenation in KH added nicorandil (0.2 μmol/L) and 5-hydroxydecanoate ( 10μmol/L) at 37 ℃.The endothelium-derived hyperpolarizing factor (EDHF) -mediated relaxation ( U46619 precontraction) induced by bradykinin in the present of indomethacin (7 μmol/L),LNNA (300 μmol/L) and oxyhemoglobin (20 μmol/L)were measured in the organ chambers.Results Compared with control group,the relaxation was significantly decreased in group A,B and D ( P ＜ 0.001 ),while there is no significant difference in group C ( P ＞ 0.05 ).Compared with group A,the relaxation was significantly reduced in group B and D (P ＜0.001 ).Conclusion Hypoxia-reoxygenation impairs EDHF mediated relaxation in coronary artery with more injury during prolonged hypoxia.This function can be restored by preconditioning with nicorandil The mechanism is mainly related to the mitochondrial ATP-sensitive K + channels.

Objective To study the effect of Perfadex and Wisconsin Unviersity solution on the function of pulmonary artery endothelium.Methods Small lobe pulmonary arteries were dissected from nine porcine lungs.The artery from each lung was cut into six rings in 2mm.Two of them were randomly incubated in Krebs,Perfadex or Wisconsin Unviersity solution (UW solution) at 4℃ for 4 hours.Endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation (percentage of-7.5 logM U46619 precontraction) induced by bradykinin or calcium ionophone A23187 in the present of indomethacin,L-NNA and oxyhemoglobin were measured at 37 ℃ in the organ chambers.Results Vasoconstriction induced by U46619 is no significant difference among three groups (P＞ 0.05).Compared with Krebs group,the relaxation induced by bradykinin or A23187 was significantly decreased in Perfadex group (P＜0.01),while there is no significant difference in UW group (P＞0.05).Conclusion Endothelium-derived hyperpolarizing factor (EDHF) plays an important role in endothelium-mediated relaxation of porcine pulmonary artery.EDHF-mediated relaxation is impaired when the lung preserved with Wisconsin Unviersity solution,wheras its function is not affected by Perfadex solution.

ObjectiveTo study the impact of type 2 diabetes mellitus on endothelium of great saphenous vein in patients with coronary heart disease.MethodsPatients undergoing coronary artery bypass grafting were selected, 20 with type 2 diabetes mellitus (experimental group) and another 20 patients without (control group).The rings of great saphenous vein in I cm length were taken from those patients and then divided into 3 segments.The structure of endothelium was evaluated by the microscope and the changes of venous tone were measured in organ chamber at 37C with a constant supply of oxygen.Venous vasoconstriction was induced by phenylephrine (10-5 mol/L) and vasodilatation induced by nitroglycerin or acetylcholine (10-9 ～ 10-5 mol/L).ResultsMore damages of ultrastructure of the endothelium of saphenous vein were found in experimental group than in control group.There were no significant differences regarding the venous tone between the two groups (P ＞0.05) when vasoconstriction induced by phenylephrine and vasodilatation by nitroglycerin.However, the vasodilatation induced by acetylcholine was significantly decreased in experimental group than in control group (P ＜ 0.05).Conclusion Type 2 diabetes mellitus can aggravate the damage of endothelium of saphenous vein in patients with coronary artery disease.

Objective To measure the expression of hypoxia-inducible factor (HIF)-1α in acral malignant melanoma (MM) tissue and to investigate its relationship with the stem cell factor (SCF)/c-kit pathway.Methods Immunohistochemical staining was performed to measure the expression of HIF-1α in tissue specimens from lesions of 93 patients with acral MM,21 with non-acral MM,39 with acral melanocytic nevi,and from the normal acral skin of 15 healthy human controls.Meanwhile,the expression of c-kit was detected by immunohistochemical staining in the 93 acral MM tissue specimens.Statistical comparisons were carried out by chi-square test and Mann-Whitney U test.The relationship of HIF-1α expression with c-kit expression as well as tumor progression and staging was assessed by Spearman correlation analysis.Results Immunohistochemistry showed that the expression rate of HIF-1α was 87.10％ (81/93) in acral MM specimens,90.48％ (19/21) in non-acral MM specimens,15.38％ (6/39) in acral melanocytic nevus specimens,but 0 (0/15) in the normal acral skin specimens.The expression of HIF-1α was significantly higher in acral MM lesions than in normal acral skin and acral melanocytic nevus lesions (both P ＜ 0.01),and significantly different between acral MM and non-acral MM lesions (P ＜ 0.01).Moreover,HIF-1α expression was positively correlated with Clark level and Breslow depth of melanoma (rs =0.442,0.368,respectively,both P ＜ 0.01),with the progression of acral MM (from in situ to aggressive and metastatic MM) (rs =0.420,P ＜ 0.01),and with the expression of c-kit (rs =0.307,P ＜ 0.01).Conclusions HIF-1α is highly expressed in acral MM,positively correlated with the staging,progression and aggression of MM,and co-expressed with c-kit in acral MM tissue,suggesting that both HIF-1α and c-kit take part in the pathogenesis of acral MM.

Molecular mechanisms underlying the effects of Fyn on cell morphology, pseudopodium movement, and cell migration were investigated. The Fyn gene was subcloned into pEGFP-N1 to produce pEGFP-N1-Fyn. Chinese hamster ovary (CHO) cells were transfected with pEGFP-N1-Fyn. The expression of Fyn mRNA and proteins was monitored by reverse transcription-PCR and Western blotting. Additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. Sequences of the recombinant plasmids pMD18-T-Fyn and pEGFP-N1-Fyn were confirmed by sequence identification using National Center for Biotechnology Information in USA, and Fyn expression was detected by RT-PCR and Western blotting. The morphology of CHO cells transfected with the recombinant vector was significantly altered. Fyn expression induced filopodia and lamellipodia formation. Based on these results, we concluded that overexpression of mouse Fyn induces the formation of filopodia and lamellipodia in CHO cells, and promotes cell movement.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Mice ; Proto-Oncogene Proteins c-fyn/genetics/*metabolism ; Pseudopodia/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time-Lapse Imaging ; Transfection

Objective To probe into the risk factors associated with posterior circulation cerebral infarction,and to evaluate the factors affecting the prognosis of posterior circulation cerebral infarction.Methods One thousand and five hundred thirty-six patients with cerebral infarction,admitted to our hospital from June 2010 to June2012,were chosen in our study; 218 patients were excluded according to the exclusion criteria; 410 patients were eliminated for incomplete information (time of onset,admission systolic blood pressure,diastolic blood pressure,glycated hemoglobin,fasting glucose,2 h postprandial glucose,previous use ofhypoglycemic agents,previous use of insulin,coronary heart disease,atrial fibrillation [AF],smoking,previous infarction or transient ischemic history of seizures,cerebral family history,triglycerides,total cholesterol,high density lipoprotein,low density lipoprotein,apolipoprotein A1,fibrinogen [FIB],and infarct type).The 908 eligible patients were divided into posterior circulation cerebral infarction group (n=431) and anterior circulation cerebral infarction group (n=477) according to their MRI features.Then the 431 patients with posterior circulation cerebral infarction were divided into possible good prognosis group (n=140) and possible poor prognosis group (n=291) according to their Barthel indexes on admission.All data were analyzed with Logistic regression analysis.Results Occurrence of posterior circulation cerebral infarction were mainly related to HbA1c≥ 8.5％ (OR=1.919,P=0.004,95％CI:1.230-2.993) and history of using insulin (OR=2.108,P=0.089,95％CI:0.892-4.982).And that of anterior circulation cerebral infarction was mainly related to AF (OR=0.655,P=0.028,95％CI:0.449-0.955).The possible prognostic factors of posterior circulation cerebral infarction were HbA1c≥ 8.5％ (OR=2.502,P=0.013,95％CI:1.217-5.147),FIB (OR=2.318,P=0.001,95％CI:1.381-3.891),time of onset (OR=2.120,P=0.011,95％CI:1.191-3.772),and age (OR=1.022,P=0.015,95％CI:1.004-1.039).Conclusion In the prevention of stroke,HbAlc can be one of the clinical biomarkers in judging stroke risk and intervention effect.

Objective To observe the relationship between the dynamic changes of Epstein-Barr virus (EBV) and the clinical efficacy of lymphoma. Methods The retrospective study was performed on 51 lymphoma patients with EBV-positive in Anhui Provincial Cancer Hospital from September 2015 to August 2017. The treatment effect of lymphoma was evaluated by the changes of EBV DNA, lactic dehydrogenase (LDH) and lymph nodes. Results Among the 51 lymphoma patients, EBV DNA elevated in 18 patients when the LDH was elevated, 23 cases presented with EBV DNA decrease and the LDH also decreased. EBV DNA decreased in 2 patients when the LDH was elevated. Eight patients were elevated when LDH was reduced. The quantification of EBV DNA and LDH had high consistency (κ = 0.609, P= 0.000). Among the 51 lymphoma patients, 20 patients with elevated EBV DNA were elevated with lymph nodeen largement, the lymph nodes in 22 cases with EBV DNA decrease were also decreased. EBV DNA decreased in 3 patients when the lymph nodes were enlarged. Six patients were elevated when lymph nodes were reduced. The quantification of EBV and lymph nodes had high consistency (κ = 0.648, P= 0.000). Patients with elevated EBV DNA during the course of treatment showed an increase in the mean value of LDH, and the patients with reduced EBV DNA during treatment also had lower average LDH. LDH levels were positively correlated with EBV DNA in patients with EBV-positive lymphoma (r= 0.627, P= 0.000). Conclusion Dynamic detection of EBV DNA and LDH can be used as the evaluation index of therapeutic effect and follow-up in patients with EBV-positive lymphoma.

Objective To explore the clinical efficacy of Da Vinci robot-assisted iliofemoral vein bypass grafting.Methods The retrospective descriptive study was conducted.The clinical data of one 66-year-old male patient who underwent Da Vinci robot-assisted iliofemoral vein bypass grafting in the First Affiliated Hospital of Army Military Medical University in March 2019 were collected.The patient was failed to recanalize iliofemoral vein stent thrombosis by endovascular measures and underwent Da Vinci robot-assisted iliofemoral vein bypass grafting after balloon occlusion preset in the common iliac vein.Observation indicators:(1) intra-and postoperative situations;(2) follow-up and survival situations.Follow-up using outpatient examination was performed to detect the patient's postoperative survival and swelling reduction of affected extremity up to April 2019.Results (1) Intra-and post-operative situations:the patient underwent Da Vinci robot-assisted iliofemoral vein bypass grafting successfully.The operation time of balloon occlusion preset by digital subtraction angiography was 35 minutes.The operation time of Da Vinci robot-assisted iliofemoral vein bypass grafting was 502 minutes (50 minutes of exposure time of femoral vein,80 minutes of exposure time of iliac vein,40 minutes of great saphenous vein harvesting time,70 minutes of end to side anastomosis between autogenous great saphenous vein and femoral vein,10 minutes of subcutaneous tunnel construction,90 minutes of end to side anastomosis between autogenous great saphenous vein and iliac vein,60 minutes of suturing except vessel closure,102 minutes of preparation time,check and washing time).The volume of intraoperative blood loss was 500 mL and no intraoperative complications occurred.The autogenous great saphenous vein graft was well filled and no bleeding was found at both proximal and distal anastomoses after iliofemoral vein bypass grafting.There were 4 abdominal Trocar holes including 2 of 1.2 cm and 2 of 0.8 cm.The incisional length of right groin and left great saphenous vein harvesting region was 5.0 cm and 15.0 cm,respectively.At the discharge time,the patient had swelling subsided partially at right lower extremity and skin tesion reduced significantly compared with the admission.The perimeters at 15 cm above right knee joint and left knee joint were 53.5 cm and 48.0 cm.The maximum perimeters of right calf and left calf were 41.0 cm and 38.0 cm.No postoperative complications occurred.Duration of hospital stay after surgery was 3 days.(2) Follow-up and survival situations:the patient was followed up for 1 month,with good survival.The patient had swelling subsided of affected extremity.The perimeters at 15 cm above right and left knee joint were 52.0 cm and 48.0 cm.The maximum perimeters of right calf and left calf were 40.0 cm and 38.0 cm.Conclusion The Da Vinci robot-assisted iliofemoral vein bypass grafting is safe and feasible,with good short-term outcomes.

Objective: To explore serum levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF), and whether changes of BDNF and GDNF are correlated with sleep quality and cognitive function in patients with chronic insomnia disorder (CID). Methods: Fifty-seven CID patients in the Department of Sleep Disorders, Chaohu Hospital of Anhui Medical University and 30 healthy controls were enrolled from May 2017 to July 2018. Pittsburgh Sleep Quality Index (PSQI) was used to assess the degree of insomnia severity (some CID patients were monitored by overnight polysomnography). Montreal Cognitive Assessment (MoCA) scale and Nine-Box Maze were used to assess general cognitive function and specific memory function, respectively. The serum levels of BDNF and GDNF were detected using ELISA. Results: Compared to the controls, CID patients had significantly higher PSQI scores (CID patients: 14.0±2.2, healthy controls: 3.9±1.1; t=28.093, P<0.01), lower MoCA scores (CID patients: 24.5±3.6, healthy controls: 26.5±0.9; t=-2.985, P<0.01), more errors in object working memory (CID patients: 1.0 (0, 1.0), healthy controls: 0 (0, 0.3)), spatial working memory (CID patients: 3.0 (2.0, 4.0), healthy controls: 1.0 (1.0, 2.0)) and object recognition memory (CID patients: 0 (0, 0), healthy controls: 0 (0, 0); Z=-2.896、-5.007、-2.306, P<0.05), and lower serum BDNF (CID patients: (19.48±7.50) ng/ml, healthy controls: (46.49±13.33) ng/ml; t=-10.274, P<0.01) and GDNF (CID patients: (32.76±14.04) pg/ml, healthy controls: (59.63±20.30) pg/ml; t=-7.240, P<0.01). The partial correlation analysis showed that in the CID patients, the levels of BDNF and GDNF were correlated with PSQI scores negatively (r=-0.293, -0.320, P<0.05) and MoCA scores positively (r=0.331, 0.295, P<0.05). The BDNF level was also correlated with the duration of disease and the errors in the spatial working memory test negatively (r=-0.319, -0.393, P<0.05), and the GDNF level was correlated with the total sleep time detected with polysomnogram positively (r=0.520, P<0.05). Conclusion Serum BDNF and GDNF levels in CID patients were lower than those in healthy controls, and correlated with sleep quality and cognitive impairment.