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Objective:To explore the pathogenesis of gallbladder cholesteryl polyps (GCP) and gallbladder cholesterol calculus (GCC) by studying the different changes of mucin (MUC) expression and reverse cholesterol transporter (RCT) in gallbladder mucosa epithelium.Methods:The data of 10 GCP patients (GCP group), 10 GCC patients (GCC group) and 5 patients with normal gallbladder resection (control group) were retrospectively analyzed, who underwent cholecystectomy in the Department of General Surgery, Xuanwu Hospital, Capital Medical University from January to December 2021. Among the 10 patients in the GCP group, there were 5 males and 5 females, aged (43.40±9.59) years old. Among the 10 patients in the GCC group, 5 males and 5 female, aged (45.00±8.13) years old. Among the 5 patients in the control group, there were 3 males and 2 females, aged (43.80±6.01) years old. Immunohistochemical analysis was used to investigate the expression differences of various subtypes of MUC and RCT [ATP binding cassette transporter G1 (ABCG1) and B group type I scavenger receptor (SR-BI)] among each group.Results:Compared with the control group, the expression of MUC1 (3.40±0.70 vs. 0), MUC5AC (1.50±0.53 vs. 0), MUC6 (4.70±0.48 vs. 0), and ABCG1 (3.50±0.53 vs. 1.60±0.55) in the gallbladder mucosa of the GCP group increased, while the expression score of SR-BI decreased (1.70±0.48 vs. 3.40±0.55), with statistical significance (all P<0.001). Compared with the control group, the expression of MUC1 (4.80±0.42 vs. 0), MUC5AC (4.70±0.48 vs. 0), MUC6 (3.30±0.67 vs. 0), and ABCG1 (3.40±0.52 vs. 1.60±0.55) in the gallbladder mucosa of the GCC group increased, while the expression score of SR-BI decreased (0 vs. 3.40±0.55), with statistically significant differences (all P<0.001). Conclusion:The different expression levels of MUC1, MUC5AC, MUC6, and RCT proteins lead to the differential formation of GCP and GCC on the basis of the co-pathogenesis in high cholesterol in bile.
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Acute appendicitis is one of the most common acute abdominal diseases in surgery. Due to its urgent onset and rapid progress, timely diagnosis and early treatment are beneficial to the prognosis of patients. The diagnosis of acute appendicitis should be based on the characteristics of different populations, based on clinical symptoms, physical examination, laboratory examination, and imaging methods. Anti-infection treatment should be started as soon as possible in the early stage, and antibiotics should choose effective drug treatment with both anaerobic bacteria and aerobic bacterial infection. Appendectomy is still the most effective treatment for acute appendicitis. In the case of uncomplicated abdominal infection such as simple and non-perforated appendicitis, antibiotic treatment may be chosen, but the development needs to be closely observed. Laparoscopic appendectomy is the first choice, but in pregnancy patients should be cautious. Emergency surgery is required for acute perforated appendicitis and attention should be paid to intraoperative abdominal flushing. Immediate surgical treatment of appendiceal abscess may involve the risk of partial ileectomy. Antibiotic treatment is recommended, with percutaneous abscess puncture and drainage if necessary.
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BACKGROUND:With the development of genetic engineering and tumor molecular biology, gene therapy for tumors has become a new treatment modality. OBJECTIVE:To explore the effect of the KDR gene silencing on the proliferation and invasion capacity of breast cancer MCF-7 cel s. METHODS:Interfering RNA (siRNA) sequences for smal molecule KDR gene was designed and transferred into human breast cancer MCF-7 cel s. Then, RT-PCR and western blot assay were used to detect the KDR mRNA and protein expression. Flow cytometry, cel counting kit-8 test and Transwel test were employed to detect the cel cycle, proliferative capacity and invasion capacity of breast cancer MCF-7 cel s after the KDR gene silencing. RESULTS AND CONCLUSION:After 48 hours of KDR silencing, the mRNA and protein expressions of KDR in MCF-7 cel s were decreased obviously;MCF-7 cel s arrested at G0/G1 stage and the number of cel s at S stage was reduced. Cel proliferation was inhibited significantly. The amount of cel s passing through the filtering membrane became less. After KDR gene silencing, the proliferation and invasion of breast cancer MCF-7 cancer stem cel s were inhibited remarkably, indicating that KDR gene silencing may be a new target for the effective treatment of breast cancer.
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Objective To research the relation of early growth response gene-1(EGR-1) and NF-κB in human T-cell leukemia virus type 1(HTLV-1) Tax protein positive cells. Methods RT-PCR was used to amplify the aimed segments EGR-1 cDNA which was then inserted into an eukaryotic expression plasmid pcDNA3.0 to construct pcDNA3.0-EGR-1. The constructed plasmid was transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, the expression levels of EGR-1, p65 and Tax mRNA in transfected cells were assay by RT-PCR after 48 h post-transfection, the proteins of EGR-1 and p65 were detected by Western blot after 48 h post-transfection too. The constructed plasmid and pNF-κB-luc reporter gene plasmid was co-transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, and the activity of luciferase was assay after 48 h post-transfection. Results The results showed that the eukaryotic expression plasmid pcDNA3.0-EGR-1 was successfully constructed. The mRNA and protein expression of EGR-1 could be promoted significantly by Tax. EGR-1 can promote the mRNA and protein expressions of p65 in TaxP cells, the activity of NF-κB was up-regulated by EGR-1 too. Conclusion EGR-1 maybe involve in adult T-cell leukemia(ATL) by increasing the activation of NF-κB.
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Objective To investigate the effect of heart preservation solution containing pinacidil on mitochondrial function in isolated rat hearts. Methods One hundred and twenty pathogen-free SD rats of both sexes weighing 250-350 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 65 mg/kg. Their hearts were immediately removed and perfused in a Langendorff apparatus. Left ventricular enddiastolic pressure was measured from a fluid-filled latex balloon inserted in the left ventricle. The isolated hearts were randomized into 5 groups (n = 24 each):group Ⅰ was perfused with cardioplegic solution HTK; group Ⅱ with HTK containing pinacidil (a non-specific sarcKATP and mitoKATP channel opener) 0.5 mmol/L; group Ⅲ with HTK containing pinacidil 0.5 mmol/L + 5-HD (a selective mitoKATP channel blocker) 100 μmol/L; group Ⅳ with HTK containing pinacidil 0.5 mmol/L + HMR-1098 100 μmol/L (a selective sarcKATP channel blocker) and group Ⅴ with HTK containing pinacidil 0.5 mmol/L + 5-HD 100 μmol/L + HMR-1098 100μmol/L. The isolated hearts were perfused with simple HTK or HTK containing pinacidil or pinacidil + 5-HD and/or HMR 20 ml/kg at 10 ml/min and then removed from Langendorff apparatus and dipped into the same HTK solution for 8 h at 4 ℃followed by 60 min reperfusion. The respiratory function of mitochondria (respiratory control rate (RCR), the rate of oxygen consumption in state 3/state 4 and P/O) was measured at the end of equilibration (T1) after 8 hpreservation (T2) and at the end of 60 min reperfusion (T3). The CK-MB and LDH activities and cTnI expression in myocardium was detected at T1 and T3. The ultrastructure of myocardium was examined at T3. Results Perfusion suspension-reperfusion (PS/R) significantly decreased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and increased myocardial cTnI concentration and CK-MB and LDH activities at T3 compared with baseline at T1 in group Ⅰ. Pinacidil significantly increased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and decreased myocardial cTnI concentration and CK-MB and LDH activities in group Ⅱ as compared with group Ⅰ-indicative of protective effect of pinacidil on mitochondria against PS/R injury. The protective effect of pinacidil against PS/R injury was attenuated by 5-HD and/or HMR1098. The myocardial damage was slightest in group Ⅱ . Conclusion Both sarcolemmal and mitochondrial KATPchannel are involved in the protective effect of pinacidil against PS/R-induced myocardial damage during heart preservation.
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Objective To evaluate the protective effect on the pulmonary micmvascolar permeability and thereby to ameliorate the lungs injury attributed to cardiopuimonary bypass(CPB).Method Twenty-four adult hybrid health dogs were randomly divided into three groups(8 in each group):group C(normal saline given after CPB),group N1(NAC given intravenously just before CPB)and group N2(NAC given just after CPB).The changes of respiratory index(RI)and malondialdehyde(MDA)content in lung tissue were observed.Samples were taken three times,before CPB(T0),30 min after CPB surned off(T1)and 60 min after CPB sumed off(T2).The leucocyte count and slbumin content in bronchoalveolar lavage fluid(BAlF),the pulmonary micmvascular permeability index(PMPI),and the histological changes of lung under light microscope and electromicroscope in 3 groups were examined.Results No significant differences were found in the levels of Ri and MDA content of lung tissue between groups before CPB.However,they gradually reduced after CPB(P<0.05)in the three groups,but they still were significantly lower in group N1 and group N2 compared with those in group C at,T1 and T2(P<0.05)and those in group N1 were significantly lowere than those in group N2 at T1 and T2(P<0.05).MDA gradually increased after CPB in three groups(P<0.05),but it was still significantly lower in group N1 and group N2 than that in group C at T1 and T2(P<0.05).The leucocyte count and albumin content in BALF were significantly lower in group N1 and group N2 in comparison with those in group C(P<0.05)and they were significantly lower in group N1 compared with those in group N2(P<0.05).The PMPI were significantly lower in group N1 and sroup N2 compared with those in group C(P<0.05)and they were significantly lower in group N1 compared with those in group N2(P<0.05).By using electromicroscope,the apparent inflammatory change of lung with endothelium cellular swelling,inter-endothelial cells spaces widened,and the indistinctness,deformation or decurtation of microfilarnent were observed.And the dissolution of laminated body,swollen mitochondria and plasmolysis were found in alveolar epithelial cell Ⅱ in group C.However,these changes were markedly alleviated in group N2 and group N1.Conclusions The results clearly demonstrate that NAC could protectie effect on the CPB injured lung and reduce the pulmonary microvascalar permeability,and the protetive effect is better in group N1 than that in group N2.
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Objective To investigate the effect of ischemic preconditioning ( IP) on the content of myocardial mitochondria cardiolipin and the heart function in isolated rat hearts. Methods Seventy-two healthy SD rats of both sexes ( 36 males,36 females) weighing 200 to 300 g were randomly divided into 4 groups ( n =18 for each group) : normal group,control group,IP group ( IP) and 5-hydroxydecanoate ( 5-HD) plus IP group ( HD) . Langendorff apparatus were used to establish the model of ischemia/reperfusion in isolated rat hearts. The hearts were perfused for 20 min to get stabilization followed by continuous perfusion in normal group for 100 min. In control group,after perfusing of 20 min for stabilization followed by continuous perfusion for 30 min,the hearts were then reperfused for 30 min after 40 min ischemia. In IP group,the hearts were given a 5-minute ischemia and 5-minute reperfusion for twice in order within a brief intermittent period,and ischemia reperfusion was carried out in the same way as that in control group. In HD group,the hearts were perfused with 100 ?mol/L 5-HD before IP,and the following procedures were carried out as those in IP group. HR,left ventricular end-diastolic pressure ( LVEDP) ,left ventricular developed pressure ( LVDP) were recorded at the end of stable perfusion ( T1) ,immediately before ischemia ( T2) and at the end of reperfusion ( T3) in all the groups. The content of myocardial mitochondria cardiolipin was measured with HPLC. Results When theparameters at T3 were compared with those at T1 and T2,HR and LVDP were decreased,LVEDP was increased and the content of myocardial mitochondria cardiolipin was decreased. All these changes were significant ( P
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AIM: To investigate the effects of the selective mitochondrial ATP-sensitive K+ channels opener diazoxide on mitochondrial respiratory function and enzyme activity in isolated rat myocardium under ischemia/reperfusion.METHODS: Observation was made on rat hearts perfused with Langendorff apparatus.72 Sprague-Dawley(SD) rats were randomly divided into 4 groups: normal group(NOR),ischemia reperfusion(IR),diazoxide group(DIA) and 5-hydroxydecanoate(5-HD) antagonized diazoxide group(5HD-DIA).Hearts isolated from SD rats were mounted on a Langendorff apparatus and started with a 20 min perfusion for equilibration.NOR went on perfusion for another 100 min after equilibration.IR underwent 40 min global ischemia and followed by 30 min reperfusion after 30 min stabilization.DIA was administered with K-H solution containing diazoxide at concentration of 50 ?mol/L for 10 min before ischemia and reperfusion.5HD-DIA was infused with 100 ?mol/L 5-HD(a specific mitochondrial ATP sensitive K+ channel blocker) and the same procedure was carried out as DIA group.Hearts were taken down to extract mitochondrial at the end-equation,before ischemia and at the end-reperfusion for determination of mitochondrial respiratory function and the enzyme activity of mitochondria.RESULTS: At the end of reperfusion,mitochondrial respiratory function(mitochondrial respiratory control rate,P/O ratio and state 3 respiration) and mitochondrial enzyme activity(NADH oxidase,succinate oxidase and cytochrome C oxidase) in DIA group were better than those in IR group and 5HD-DIA group(P0.05).CONCLUSION: Preconditioning with mitochondrial ATP sensitive potassium channel opener,diazoxide,protects rat heart mitochondria against ischemia-reperfusion injury.The mechanisms are involved in the safeguarding of respiratory function and activity of enzymes of respiratory chain.
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Objective To compare the myocardial protective effect of hyperpolarized cardioplegic arrest with pinacidil against that of depolarized hyperkalemic arrest during cardiopulmonary bypass Methods Eighteen healthy mongrel dogs of both sexes weighing 10 15kg were divided into three groups of six each, according to the cardioplegic solution infused into aortic root after aortic cross clamp was applied; group A received 4℃ standard St Thomas solution (K +16mmol/L); group B 37℃ St Thomas solution (K +5mmol/L) containing pinacidil (50?mol/L) mixed with blood (the ratio was 1:1);group C 4℃St Thomas solution (K + 5mmol/L) containing pinacidil(50?mol/L) mixed with blood (1∶1) The global ischemic time was 60min followed by 30min reperfusion Myocardial tissue was taken before and 30min, 60min after aortic cross clamping and 30min after reperfusion for determination of myocardial content of adenine nucleotide and MDA and ultrastructure examination with electron microscope Hemodynamics (BP,CVP,PCWP,CO,CI and LVSW) was also measured before aortic cross clamping and 15,30min after declamping Results Significant ischemic and reperfusion damages were found in group A, while there were only mild damages in group B and C Hemodynamics recovery after aortic declamping was significantly better in group C than that in group A and B Conclusions Myocardial protective effect of hyperpolarized arrest produced by blood cardioplegic solution containing pinacidil is superior to that of traditional depolarized hyperkalemic arrest and hypothermic hyperpolarized cardioplegia is better than normothermic
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The myocardial protective effect of continuous infusion with normothermic oxygenated crystalloid solution or blood cardioplegia during normothermic CPB was studied in 15 dogs. Ultrastructure. levels of adenine nucleotides,lipid peroxide (LPO),water content of heart musle,hemodynamics were observed. Dogs were randomly divided into three groups. Group Ⅰ: intermittent infusion with cold crystsalloid cardioplegia during hypothermic CPB (n=5); Group Ⅱ: continuous infusion with normothermic oxygenated blood cardioplegia during normothermic CPB(n=5); Group Ⅲ: continuous infusion with normothermic oxygenatde crystalloid cardioplegia duriug normothermic CPB((n=5). The normal mitochandria contents and glycogen stores in group Ⅱ and Ⅲ were significantly higher than those in group Ⅰ(P
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Objective To compare the myocardial protective effect of pinacidil-induced hyperpolarized and hyperkalemic depolarized cardiac arrest on isolated rabbit heart under different temperature. Methods Forty-eight rabbits of both sexes weighing 2.0-2.5 kg were sacrificed by a knock on the head. Their hearts were excised and perfused in a Langendorff apparatus with an oxygenated Krebs-Hensleit buffer (KHB)(37℃). The study was divided into six groups: A normothermic hyperkalemic cardioplegia; B normothermic hyperkalemic blood cardioplegia; C hypothermic hyperkalemic cardioplegia; D hypothermia hyperkalemic blood cardioplegia; E normothermic hyperpolarizing blood cardioplegia; F hypothermic hyperpolarizing blood cardioplegia. Three pieces of myocardial tissue were obtained from apex of left ventricle at the end of the study for determination of myocardial adenine nucleotide and lipid peroxide content and microscopic examination. The following were recorded : (1) cardiac arrest time: the time from perfusion with cardioplegic solution to the beginning of cardiac arrest. (2) heart beat recovery time ;the time from reperfusion with KHB to the beginning of normal heart beat. (3) changes in HR, left ventricle developed pressure and myocardial contractility before and after cardiac arrest. Results The cardiac arrest time was longer and the time for the heart to restart was shorter in the two hyperpolarizing blood cardioplegia groups (group E and F) than that in the other 4 groups. No arrhythmia occurred in group E and F. Left ventricle developed pressure(LVDP) and left ventricle contractility recovered quickly after reperfusion with KHB was started and were restored to the pre-ischemia level after 20 min in group E and F. The levels of ATP, TAN and EC were higher and the MDA level was lower in group E and F than those in the other 4 groups. Myocardial structure was less injuried in group E and F. Conclusion The myocardial protection effect of hyperpolarizing blood cardioplegia with pinacidil is superior to traditional hyperkalemic depolarizing cardioplegia.
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ve To assess the protective effect of L-arginine on vascular endothelial cell during cardiopulmonary bypass(CPB). Methods 12 adult healthy mongrel dogs of either sex weighing 10-15kg, were divided into L-arginine group (group L, n=6) and control group (group C, n=6) . Femoral artery and vein were cannulated for monitoring of MAP and CVP. Right internal jugular vein was cannulated with Swan-Ganz catheter for hemodynamic monitoring. L-arginine 100 mg?kg-1 was administered intravenously before CPB followed by continuous intravenous infusion at 10mg?kg-1.min-1 to the end of CPB in group L. Control group was treated as group L except the administration of L-arginine. Venous blood samples were taken before CPB, 5min after CPB was started, 15, 45min after aortic cross-clamping and 15, 30min after release of aortic cross-clamp for determination of the plasma levels of metabolites of nitric oxide (NO2- and NO3-), malondialdehyde (MDA) and von Willebrand Factor(vWF). Hemodynamic parameters were measured before CPB, 15 and 30min after release of aortic clamp. Specimens of tissue from femoral artery and descending aorta were taken for microscopic examination at 15,45 min after aortic cross-clamping and 30min after release of aortic cross-clamp.Results There was little difference in plasma NO2- and NO3-concentrations before CPB between the two groups. NO2- and NO3- levels increased significantly after CPB in both groups but were significantly higher in group L than those in group C (P
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Objective To evaluate the myocardial protective effect of hyperpolarized arrest induced with ATP-sensitive potassium channel opener ,pinacidil in vitroMethods Twenty-four rabbit hearts were randomly divided into three groups : hypothermic hyperpolarized group (LH group), normothermic hyperpolarized group ( WH group), and hyperkalemic control group (group C) The isolated rabbit hearts with a Langendorff apparatus were perfused oxygenated Krebs-Henseleit's solution (K-H) for 10 min and followed by cardioplegia The cardioplegia consisted of StThomas solution with either traditional high potassium (16mmol/L KCl, 4 ℃, group C), or pinacidil 50?mol/L (4 ℃ in LH group or 37 ℃ in WH group ) with 5mmol/L KCl The hearts were subjected to 40 min perfusional occlusion and followed by 20 min reperfusionResults Hearts were arrested quickly in LH group, and a little slowly in WH group, but rebeated quickly , which were different significantly from those in group C ; Recovery of LVP and left ventricle contracility in LH group and WH group were remarkedly faster than those in group C (P
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Objective To evaluate the myocardial protective effect of hyperpolarized heart arrest at different temperature during cardiopulmonary bypass CPB.Methods Eighteen dogs were randomly allocated to be infused with St.Thomas solution containing 50?mol/L pinacidil and 5mmol/L K + at 37℃ (group A) or 4℃ (group B), or the standard St.Thomas solution containing K + 16mmol/L at 4℃ (group C) respectively, through aortic root after aortic clamping . The global surgical ischemia lasted 60min with the declamping of 20min .The activities of serum myocardial enzymes, and levels of lipid peroxide and adenine nucleotide of myocardium were measured.Results All paramaters showed to occur serious ischemic and reperfusion damages during the whole procedures in group C, while there were the mild damages in two hyperpolarized groups, especially in group B.Conclusions Myocardial protective effect of hyperpolarized arrest induced with pinacidil, an ATP sensitive potassium channel opener, is superior to that of traditional hyperkalemic cardioplegia , which is much more prominent in hypothermic state.
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Objective: To study the protective effect of coenzyme Q_10 on red blood cell membrane and its immunity in patient during extracorporeal circulation. Method: Twenty patients under extracorporeal circulation about 30 minutes were randomly divided into control group and coenzyme Q_10 group (n=10). Coenzyme Q_10 2mg/kg was added to the priming fluid in coenzyme Q_10 group. Plasma levels of malondialdehyde (MDA), free hemoglobin (FHb), immune adhe sion ability and immune compound of red blood cell membrane were measured before extracorporeal circulation, 15 and 30 minutes after beginning of cardiopulmonary bypass, and on first day morning after operation. Result; All different periods after beginning of cardiopulmonary bypass in coenzyme Q_10, group, MDA and FHb levels were lower and im mune adhesion ability was higher than those of control group. Conclusion: Coenzyme Q_10 may protect red blood cell membrane and its immunity of patient during extracorporeal circulation.