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To explore whether the downregulation of protein phosphatase 2A catalytic subunit(PP2Ac)involved in the pathogenesis of mitochondria fission/fusion dynamics and functional imbalance induced by human tau accumulation. After cotransfection with mito-dsRed plasmids and pIRES-eGFP-tau40 plasmids 48 hours,the rat primary hippocampal neurons were observed with a laser scanning confocal microscope for their changes in shape and distribution of mitochondria.The expressions of mitochondria fission/fusion protein and PP2Ac and PP2Ab were detected by Western blotting.Furthermore,the shape and distribution of mitochondria of rat primary hippocampal neuron and wild type 293wt cells were assayed 48 hours after co-transfection with siPP2Ac-EGFP plasmids and mito-DsRed plasmids,and the fission/fusion dynamics of 293wt cells was captured with live cell time-lapse imaging after co-transfection with siPP2Ac plasmids and mito-Dendra2 plasmids.After transfection with siPP2Ac plasmids,the relative level of mitochondria fission/fusion protein of 293wt cells was assayed by Western blotting,and mitochondria membrane potential was detected by JC-1 staining,and the cellular viability was measured by CCK8 assay.Finally,the shape and distribution and membrane potential of mitochondria of HEK293 cells with stable transfection of htau40(293htau)were detected after co-transfection with PP2Ac and mito-dsRed plasmids. Human tau40 expression decreased distribution of mitochondria and significantly lowered PP2Ac level in primary hippocampal neuron(=4.814, =0.0086).Down-regulation of PP2Ac caused mitochondria elongation and perinuclear accumulation in primary hippocampal neuron and 293wt cells;in addition,down-regulation of PP2Ac in 293wt cells significantly increased mitochondria fusion rate(=2.857, =0.0074)and the levels of mitochondria fusion protein mitofusin(MFN)1(=6.768, =0.0025),MFN2(=3.121, =0.0035),and optic atrophy 1(=3.775, =0.0199);however,the levels of dynamin-like protein-1 and Fis1 remained unchanged.The down-regulation of PP2Ac in 293wt cells led to the significant decrease in mitochondria membrane potential(=2.300, =0.0270)and cell viability(=6.249, <0.0001).Finally,up-regulation of PP2Ac attenuated the abnormalities in the shape,distribution and function of mitochondria in the 293htau cells. Down-regulation of PP2Ac is involved in the abnormal shape and distribution of mitochondria and its dysfunction induced by human tau40 in rat primary hippocampal neurons and HEK293 cells.
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Animais , Humanos , Ratos , Domínio Catalítico , Regulação para Baixo , Células HEK293 , Mitocôndrias , Proteína Fosfatase 2 , Proteínas tauRESUMO
Objective To evaluate the effect of medical hygiene wipes on disinfecting object surface and compliance to use in an intensive care unit (ICU).Methods 30 beds in the ICU at a hospital were randomly selected and divided into three groups:A,B and C,each group had 10 beds.Group A were only provided hygiene wipes,group B used towel soaked in chlorine-containing disinfectant (disinfected towel),group C were provided both medical hygiene wipes and disinfected towel for selected use.Compliance to different disinfection methods,and disinfectant efficacy on object surface were compared.Results The rate of compliance to disinfection in group A was higher than group B (78.78% vs 42.12%,χ2 =68.56,P <0.05);except medical disposal trolley and ventilator,compliance to disinfection of the other object surface in group A were all higher than group B (all P <0.05).Except ventilator, adenosis triphosphate (ATP)values before and after disinfection of object surface in group A and B were both significantly different (both P <0.05 ),disinfectant efficacy of group A was superior to group B.Group C used medical hygiene wipes more than disinfected towel ([836 times,91 .57%]vs [77 times,8.43%]).Nurses spent (20.15+ 3.00 )minutes on preparing disinfectant every day,and (15.02 ± 2.00 )minutes on washing used disinfected towel,while hygiene wipes could be used immediately,discarded after use,and was timesaving. Conclusion Use of medical hygiene wipes is convenient,rapid,and without irritating odor,it also improves disinfection effect on object surface and ICU health care workers’compliance to disinfection.
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OBJECTIVE:To discuss the application practice of PIVAS standardization in pharmacy intravenous admixture ser-vice(PIVAS)of our hospital. METHODS:Based on JCI,the design concept and management mode of PIVAS were analyzed and summarized from PIVAS design,management,job and medical order time arrangement,staff management,infusion quality or fault emergency plan,etc. RESULTS:After introducing JCI standards,the improvement was conducted in respects of management system and work flow design,purification system and logistics division design,information system design,etc. Working hour was arranged according to the workload or work flow and the actual needs of clinical departments. Medical orders,1 batch per hour,24 batches one day,were delivered to clinical departments according the time point. The positivity of the staff had been improved, and work quantization and performance appraisal had been achieved. Those improvement optimized drug dispensing process, cleared the responsibility of information management and guaranteed the quality of infusion. Work error analysis and summary were monthly conducted;the number of work error decreased from 562 cases before JCI introduction(2013)to 264 cases after JCI intro-duction (2014),decreasing by 112.87% annually. CONCLUSIONS:JCI standard is of significance to the promotion of PIVAS management,work level and quality,and safe and rational use of drugs in the clinic.
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AIM:To investigate the protective effects of astrocyte protein phosphatase 2A (PP2A) up-regula-tion on APP/PS1 double transgenic mice.METHODS:An eGFP-wtPP2A lentivirus with glial fiber acidic protein promoter was constructed to specifically increase PP2A expression in the astrocytes.The mice were divided into wild -type mice +vector virus group (Con), APP/PS1 transgenic mice +vector virus group (APP/PS1) and APP/PS1 transgenic mice +eGFP-wtPP2A lentivirus group (PP2A) by lateral ventricular injection of the lentivirus.Four weeks after injection of the vi-rus, the immunofluorescence of brain slices were used to detect the level of β-amyloid protein ( Aβ) .Golgi staining was used to detect the changes of dendritic spine density and morphology.Electron microscopy was applied to detect the thickness of postsynaptic density (PSD).The Morris water maze test was applied to examine the learning and memory abilities of the mice.RESULTS: Up-regulation of PP2A in the astrocytes attenuated Aβlevel increasing in APP/PS1 group.Up-regulation of PP2A in the astrocytes significantly attenuated both decreases in the dendritic spine density and the percentage of mushroom-like dendritic spines in the hippocampal CA3 region of APP/PS1 mice.Up-regulation of PP2A in the astrocytes significantly attenuated the reduced thickness of PSD in APP/PS1 group.Up-regulation of PP2A in the astro-cytes attenuated the escape latency extending in APP/PS1 group .CONCLUSION: Up-regulation of PP2A in the astro-cytes reduces AD-like pathological changes, and attenuates synaptic impairment, synaptic plasticity deficits and cognitive impairment in the APP/PS1 double transgenic mice.
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〔Abstract〕 The paper introduces the informatization management system in specialist clinic blood-taking room, which compares with the process before informatization from the aspects of error rates of blood samples, patients satisfaction, hospital labor costs and so on to highlight the advantages of informatization management.The progressiveness, existing problems and corresponding countermeasures are discussed, informatization management could improve work efficiency and patients satisfaction.
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<p><b>OBJECTIVE</b>To identify the genetic cause for a Chinese Han family affected with primary hypertrophic osteoarthropathy.</p><p><b>METHODS</b>Whole blood and urine samples were collected from a patient and 7 unaffected relatives of the family. The coding sequences and intron/exon boundaries of HPGD and SLCO2A1 genes of the patient were amplified with polymerase chain reaction and sequenced. The genotypes of relatives were subsequently verified. Urinary prostaglandin level was measured with enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>A homozygous 2-bp deletion in HPGD gene (c.310_311delCT, or p.L104AfsX3) was detected in the patient, and 5 heterozygous carriers were identified in the relatives. The urinary prostaglandin E2 (PGE2) level was significantly elevated (P<0.01), while PGE-M was significantly reduced (P<0.01) in the patient.</p><p><b>CONCLUSION</b>Primary hypertrophic osteoarthropathy in this family is caused by a homozygous mutation (c.310_311delCT) in the HPGD gene.</p>
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Adulto , Feminino , Humanos , Masculino , Povo Asiático , Etnologia , Genética , Sequência de Bases , Dinoprostona , Urina , Hidroxiprostaglandina Desidrogenases , Genética , Dados de Sequência Molecular , Mutação , Transportadores de Ânions Orgânicos , Genética , Osteoartropatia Hipertrófica Primária , Diagnóstico , Etnologia , Genética , LinhagemRESUMO
Objective To investigate the difference of CCAAT/enhancer-binding protein α (C/EBP-α) gene induced apoptosis between hepatocytes and hepatic stellate cells (HSC) in mice with liver fibrosis.Methods Sixty BALB/c mice were evenly divided into normal group,model group,treatment group,blank control group and negative control group,12 mice in each group.Except the mice of normal control group,the mice of other groups were treated with intraperitoneal injection of CCl4 to establish liver fibrosis mice model.Mice of treatment group,blank control group and negative control group were administrated with C/EBP-α carried adenovirus (Ad-C/EBP-α),phosphate buffered solution and empty vector of adenovirus (Ad-EGFP) respectively through tail vein for the first week.The expression of C/EBP-α and α-smooth muscle actin (α-SMA) was detected by immunohistochemistry method.Sinusoidal endothelial structure of peri-portal regions and far from portal regions was observed by transmission electron microscope (TEM).Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was applied to detect apoptosis of cells in liver tissue.The degree of liver fibrosis in mice was determined with sirius red staining and hydroxyproline content measurement.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between two groups.Results C/EBP-α was expressed in nucleus of hepatocyte in normal control group mice.The expression decreased in model group,blank control group and negative control group.However,the expression of C/EBP-α of treatment group increased,and mainly expressed in cells located in perisinusoidal and perivascular.Hepatic sinusoids was distorted,blood vessel wall thickened.Hepatocyte degeneration and lots of lipid droplets was found in model group,blank control group and negative control group.The thicken degree of endothelial layer of blood vessel of treatment group was lower than that of model group.The percentage of sirius red positive cells of normal group,model group,treatment group,blank control group and negative control group was (0.10±0.03)%,(5.81±0.32)%,(2.32±0.45)%,(6.34± 0.81)% and (6.10± 0.92)%,respectively; content of hydroxyproline was (0.07±0.00) μg/mg,(0.69 ± 0.10) μg/mg,(0.19±0.06) μg/mg,(0.56±0.03) μg/mg and (0.64±0.08) μg/mg,respectively; the percentage of α-SMA positive cells was (0.50 ±0.03)%,(5.30 ± 0.52)%,(2.15 ± 0.29)%,(5.53 ± 0.43) % and (5.42 ± 0.25) %,respectively; the number of TUNEL positive cells was (0.25 ± 0.08),(0.15±0.02),(7.10±1.53),(0.13±0.03) and (0.18±0.07),respectively.The differences between the groups were statistically significant (F=113.74,148.29,292.43 and 140.25,all P<0.05).The difference between normal group and model group,between model group and treatment group,between treatment group and blank control group,between treatment group and negative control group were statistically significant (tarirus positive cell =-52.54,-16.20,-10.60 and-7.99,thydroxyproline content =-168.00,11.53,11.07 and 12.54,ta SMA pusitive cells-24.77,-13.82,15.94 and 18.37,tTUNEL positive cells =3.26,-11.91,-11.95 and-11.88,all P< 0.05),there was no statistically significant difference between model group and blank control group,between model group and negative control group (both P>0.05).TUNEL positive cells mainly located in perisinusoidal and perivascular of liver in mice,which was consistent with the distribution of α-SMA-positive cells.Conclusion C/EBP-α could effectively relieve CCl4 induced liver fibrosis in mice mainly through inducing HSC apoptosis,however no apoptosis effect on hepatocytes.
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Objective To study the anti-proliferation and apoptosis induction effect of cisplatin combined with Astragalus on human laryngeal cancer Hep-2 cells.Methods The proliferation inhibition of cisplatin and Astragalus alone or in combination on Laryngeal cancer Hep-2cells was measured by MTT assay.Effects of cisplatin and Astragalus alone or in combination on apoptosis of Hep-2 cells were detected by flow cytometry (FCM).Western blot was used to analyze the expressions of Bcl-2 and Bax proteins.Results The inhibition ratio of proliferation of Hep-2 cells was (53.83 ± 17.12) % in the astragalus group,(69.12 ± 27.12)% in the cisplatin group,and (84.55 ± 27.84)% in the cisplatin combined with Astragalus group,and was significantly greater than the control group (0%) (t =16.87,16.67,40.90,P <0.01),respectively.The apoptotic ratio of Hep-2 cells was (38.2 ± 13.6)% in the astragalus group,(67.2 ± 17.8)% in the cisplatin group,and (86.4 ± 25.1)%] in the cisplatin combined with Astragalus group,and was significantly greater than control group (17.1 ± 1.3) % (t =8.11,12.77,24.92,P <0.05),respectively.The effect in the combination group is better than the other group (t =11.33,9.37,P < 0.01).The expressions of Bcl-2 and Bax proteins were changed.Conclusions The tumor-killing effect of cisplatin on laryngeal cancer Hep-2 cells could be enhanced significantly by the combination application of astragalus by the way of regulating the expression of Bcl-2/Bax.
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Objective To study the tumorigenetic characteristics of side population (SP) cells and their exsistence in gastric cancer cell lines and tissues.Methods Flow cytometry and the DNA-binding dye Hoechst 33342 staining were used to analyze sorted SP cells in 3 gastric cancer cell lines(SGC-7901,BCIC-823 and MKN28).SP cells and non-SP cells(main population cells)isolated from SGC-7901 cell line weresubcutaneously injected into 36 nude mice with 500,5000 or 50 000 cells per mouse,respectively.Thetumorigenity was observed 8 weeks after injection.The expression of ATP-binding cassette sub-family Gmember 2(ABcG2)mRNA in three gastric cell lines and its level in gastric cancer tissues were detected by real-time PCR and immunostaining.respectively.Results The proportion of SP cells was accounted for 1.0%in SGC-7901 cells and 1.3%in BGc-823 cells.No SP cell was found in MKN28 cells.As low as 500 SP cells or 50 000 non-SP cells could initiate tumors in mice.The expression of ABcG2 mRNA was higher in SGC-7901(O.162)and BGC-823(O.096)cell lines than that in MKN28 cell line(0.005).The value of AtCG2 mRNA/beta-aetin mRNA and protein in gastric cancer tissues was different from those in gastritis tissues.Condasimm SP cells have strong ability of tumorigenesis compared with non-SP cells.The expression of ABCG2 is found in gastric cancer tissues and part gastritis tissues.The more the SP cells in gastric cell lines,the higher the expression of ABCG2.
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Human interleukin-15 (hIL-15) is an important cytokine to activate endothelial cells and can be regulated by many other cytokines. The aim of this study is to examine the ability of interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) to induce the production of human interleukin-15 (hIL-15) and IL-15 receptor (IL-15Ralpha) by human umbilical vein endothelial cells (HUVECs). The data are summarized as follows: 1. Northern blot revealed that IL-15 mRNA was up-regulated by IFN-gamma and TNF-alpha. 2. Intracellular IL-15 protein was visualized by fluorescence microscopy, whereas the expression of IL-15 on the surface of HUVECs was detected by fluorescence activated cell sorting (FACS), and no detectable IL-15 in the medium was verified by ELISA. 3. IL-15Ralpha was detected on the surface of HUVECs by FACS after IFN-gamma and TNF-alpha stimulation, whereas Western blotting revealed that the elevated expression on surface IL-15Ralpha was not due to the increased protein expression. The conclusion demonstrated from our results is that IFN-gamma and TNF-alpha play an important role in regulating the expression of IL-15 and IL-15Ralpha on the surface of HUVECs.
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OBJECTIVE: To investigate the utilization and its trend of antidepressants in Beijing.METHODS: The antidepressants used in 70 hospitals in Beijing from 2004 to 2006 were analyzed statistically in respect of the variety,consumption sum,DDDs and DDC etc.RESULTS: The consumption sum of the antidepressants used in Beijing increased year by year.CONCLUSION: Selective serotonin reuptake inhibitors(SSRIs) have become the first-line antidepressants,but the application of the tricyclic and tetracyclic antidepressants decreased year by year.The new antidepressants such as venlafaxine increased sharply.
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Human interleukin-15 (IL-15) is a proinflammatory cytokine to suppress neutrophil apoptosis, which is a potential therapeutic agent. The modulatory effect of TNFα was investigated in IL-15-induced suppression of human neutrophil apoptosis. TNFα was shown to reverse the ability of IL-15 to delay neutrophil apoptosis within certain time course. Moreover, this reverse effect by TNFα might be associated with a reduction of the expression of the anti-apoptotic Bcl-Xl protein detected by Western blotting. It is concluded that TNFα can be used to modulate IL-15-induced suppression of neutrophil apoptosis within certain time course.
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Objective The study aimed to know the mental health state of juvenile delinquents, to provide basis for the early prevention of juvenile delinquency and the rectification of the unhealthy mental state. Methods The 378 juvenile delinquents from Tianjin reformatory were deem to the study group, the 410 14~18 years old high school students were sampled from a common school as the control group. They were assessed and compared with SCL-90. Result The indices of SCL-90 excluding somatic change, interpersonal relationship were higher than that of the domestic routine model (P
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AIM: To study the pharmacodynamics of Yajiao Hadun San(Vitex trifolia;streptocaulon griffibhii;Asparagus pseudofilicinus;Inula cappa;Benincasa hispida;Tacca esquirolli). METHODS: The pyrexia model rat induced with active yeast,the mice's auricular swelling test caused by croton oil;the mice's toe swelling test caused by albumen;the mice's twisting body test caused by the irritation of acetic acid and the mice's carbon clearance test were used to determine antipyretic,analgesic,anti-inflammation effect and immunity function of Yajiao Hadun San. RESULTS: Yajiao Hadun San could reduce the anus temperature of the pyrexia model rat,inhibit the mice's toe and auricular swelling,reduce the times of twisting body sharply and also increase the rate of eliminating colloid carbon in mice. CONCLUSION: Yajiao Hadun San has certain antipyretic,anti-inflammation,analgesic effect and enhance the phagocytosis of mono-karyon macrophage in mice.
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@#Objective To observe the effect of morroniside on cerebral infarction volume in focal cerebral ischemia-reperfusion rat model.Methods The animal model was induced by occlusion of middle cerebral artery with suture embolus, ischemia for 30 minutes, and reperfusion for 7 days. The infarction volume was measured by 2,3,5-triphenyltetrazolium chloride(TTC) staining technique.Results Compared with sham operation group, the cerebral infarction volume ratios increased obviously in model group and in the drug-treated(90 mg/kg,270 mg/kg)groups. Compared with model group, the cerebral infarction volume ratios decreased obviously in the morroniside-treated(90 mg/kg,270 mg/kg)group,while the cerebral infarction volume ratios in vitamin E-treated(35 mg/kg)group didn't change.Conclusion Morroniside may decrease the cerebral infarction volume after cerebral ischemia-reperfusion. It possesses protective effect against cerebral ischemia-reperfusion injury.
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@#The cerebral ischemia reperfusion injury is one of factors in aggravating brain injury, and the inflammatory response is one of the main reasons in the reperfusion injury after acute cerebral ischemia. Inflammation is modulated by many factors such as inflammatory mediators and inflammatory cells that promote brain damage from ischemia injury to reperfusion injury. This paper would review the role of inflammatory cells and mediators such as cytokines, chemotactic factors, adhesion molecules on cerebral tissue injury.
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AIM: To study the effects of 1.5 MAC halothane and sevoflurane on ischemic myocardium. METHODS: The isolated rat heart were perfused with halothane and sevoflurane and HR, LVEDP, LVDP, +d p /d t , -d p /d t , coronary flow (CF), the myocardial ATP content and Ca 2+ -ATPase activity were determined before and 10 min and 25 min after ischemia. In the meantime, LVP was recorded during 25 min ischemia. RESULTS: 1.5MAC sevoflurane significantly increased CF in normal isolated rat hearts. Both halothane and sevoflurane depressed myocardial contractile function, increased normal myocardial energy storage. After 10 min ischemia, the decrease of myocardial ATP content were slowed down by halothane and sevoflurane, especially halothane. During 25 min of ischemia, the onset time of contracture was significantly delayed, and the contracture intensity was alleviated by halothane, but not sevoflurane. CONCLUSION: Halothane has better protective effect on ischemic myocardium than sevoflurane through preventing the decrease of myocardial ATP content and Ca 2+ -ATPase activity during ischemia.