α-硫辛酸对高糖作用下人脐静脉内皮细胞护骨素表达的影响

目的 观察α-硫辛酸对高糖作用下血管内皮细胞护骨素(OPG)mRNA表达的影响.方法 脐静脉内皮细胞株分别在5.5mmol/L葡萄糖、30mmol/L葡萄糖及30mmol/L葡萄糖+不同浓度α-硫辛酸(50、100、200μmol/L)条件下培养48h.取内皮细胞培养上清液,以黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,以硫代巴比妥酸为底物检测丙二醛(MDA)含量;RT-PCR法检测内皮细胞OPG mRNA表达水平的变化.结果 高糖作用下内皮细胞SOD活性降低(P<0.01),MDA水平升高(P<0.01),OPG mRNA表达水平显著上升(P<0.01);以不同浓度α-硫辛酸干预高糖组,SOD活性升高(P<0.01),MDA水平降低(P<0.01),OPG mRNA表达水平显著下降(P<0.05).结论 α-硫辛酸改善高糖环境下血管内皮细胞的氧化应激状态,下调OPG mRNA的表达水平.     

α-硫辛酸对糖基化终产物诱导血管内皮细胞凋亡的影响

目的观察α-硫辛酸对糖基化终产物诱导血管内皮细胞凋亡的影响。方法体外培养人脐静脉内皮细胞,以人血清清蛋白作为阴性对照,用含不同浓度梯度(100、200、400mg/L)糖基化终产物的培养液对内皮细胞体外培养60min及α-硫辛酸200μg/ml作用30min后再加入200mg/L糖基化终产物作用内皮细胞60min,采用ELISA法检测内皮细胞8-OHdG水平,瑞氏-吉姆萨染色及Hoechst33258染色观察内皮细胞形态学变化。结果糖基化终产物组内皮细胞8-OHdG水平较对照组明显增加(P0.05),糖基化终产物以剂量依赖方式诱导培养的内皮细胞凋亡,α-硫辛酸干预后内皮细胞8-OHdG水平显著降低,明显减轻内皮细胞凋亡的形态学改变。结论α-硫辛酸抑制糖基化终产物诱导内皮细胞凋亡,其作用机制可能是通过降低细胞氧化应激水平。     

Protective effect of osteoprotegerin in vascular endothelial cells cultured with high glucose and its possible mechanism

目的 探讨护骨素对高糖诱导的血管内皮细胞保护作用及机制.方法 人脐静脉内皮细胞分别在正糖(5 mmol/L葡萄糖)、高糖(25 mmol/L葡萄糖)、高糖+护骨素(25 mmol/L葡萄糖+2 μg/ml护骨素)、高渗(5mmol/L葡萄糖+20 mmol/L甘露醇)环境下培养72小时.以流式细胞术测定各组细胞的凋亡;Western blot法分析各组细胞磷酸化核转录因子(NF-κB)抑制蛋白激酶β (p-IκKβ)、NF-κB抑制蛋白激酶β(IκKβ)、NF-κB抑制蛋白α(IκBα)、bcl-2及bax表达水平.结果 (1)与正糖组相比,高糖组内皮细胞凋亡率显著增加(P＜0.01);高糖+护骨素组细胞凋亡明显低于高糖组,而又高于正糖组(P＜0.05);(2)与正糖组相比,高糖组p-IκKβ表达显著增高(P＜0.01),IκBo和bcl-2表达显著降低(P＜0.01);高糖+护骨素组p-IκKβ明显低于高糖组,而又高于正糖组(P＜0.05),IκBα和Bcl-2表达水平明显高于高糖组,而又低于正糖组(P＜0.05).结论 高糖通过激活IκKβ/NF-κB信号通路引起血管内皮细胞凋亡;护骨素具有抗高糖诱导的IκKβ/NF-κB活性作用,从而降低内皮细胞凋亡,保护内皮细胞.     

艾塞那肽抑制高糖诱导内皮细胞凋亡的作用机制

在正常糖浓度(5.5 mmol/L)和高糖(33 mmol/L)培养的人脐静脉内皮细胞(HUVECs)分别加入不同浓度的艾塞那肽并干预不同时间后,噻唑蓝(MTF)法检测细胞活力,应用流式细胞仪检测细胞早期凋亡率,Western印迹法检测蛋白激酶B(Akt)磷酸化、Bcl-2和Bax蛋白表达水平.结果显示,HUVECs经高糖培养48和72 h后,细胞活力明显下降(P＜0.01).经1、10和100 nmol/L艾塞那肽干预48 h后,细胞活力明显增加,并呈浓度依赖性(P＜0.01).与正常糖浓度组比较,高糖组细胞凋亡率升高,Akt磷酸化和Bcl-2蛋白表达水平下降,Bax蛋白表达增加,Bcl-2/Bax比值降低(P＜0.01).艾塞那肽干预后,细胞凋亡率下降,Akt磷酸化和Bcl-2蛋白表达增加,Bax蛋白表达下降,Bcl-2/Bax比值升高(P＜0.01),而艾塞那肽的作用可被磷酸肌醇3激酶(PI3K)抑制剂LY294002所对抗(P＜0.01).提示艾塞那肽可通过PI3 k/Akt信号通路调节Bcl-2/Bax蛋白表达来抑制高糖诱导的内皮细胞凋亡,起到保护内皮细胞的作用.     

丹皮酚对高血压患者血管内皮细胞Toll样受体4、核因子-κB表达的影响

目的 观察丹皮酚对Toll样受体(TLR)、核因子(NF) -κB信号转导通路的影响,探讨其对高血压患者血管内皮细胞的保护机制.方法 10％高血压患者及健康人血清干预人脐静脉内皮细胞(HUVEC-C)24 h,实时荧光定量(PCR)法检测TLR4 mRNA表达.不同浓度的丹皮酚干预脂多糖(LPS)诱导的HUVEC-C TLR4表达(24 h)及NF-κB活化(2 h),PCR法测TLR4 mRNA及NF-κB mRNA的表达,免疫印迹技术测TLR4蛋白及NF-κB蛋白的表达.结果 血清作用24 h后,高血压病组TLR4 mRNA的表达较健康组增高(P＜0.01).丹皮酚可呈剂量依赖性减少LPS诱导的TLR4 mRNA高表达,抑制LPS诱导的TLR4蛋白高表达和IκBα蛋白的降解(P＜0.05,P＜0.01,P＜0.001).结论 TLR-NF-κB信号途径介导的炎症反应和免疫紊乱是高血压病血管内皮损伤的机制之一,丹皮酚可通过抑制其表达保护血管内皮细胞损伤.     

N-乙酰半胱氨酸通过抑制JAK2/STAT3信号通路对高糖诱导的内皮细胞损伤的保护作用

目的 探讨N-乙酰半胱氨酸(NAC)能否通过抑制JAK2/STAT3信号通路对高糖诱导的内皮细胞损伤起保护作用。方法 体外分别应用不同浓度NAC对人脐静脉内皮细胞(HUVEC)进行预处理,筛选出NAC减轻高糖诱导的HUVEC细胞毒性的最佳浓度。使用最佳浓度的NAC预处理高糖诱导的HUVEC,通过CCK-8检测细胞存活率,Hoechst33258核染色荧光显微镜照相法检测内皮细胞凋亡的形态学改变,Western blot检测JAK2/STAT3信号通路的蛋白表达水平,DCFH-DA检测细胞内活性氧水平,ELISA检测相关炎症因子细胞间黏附分子1(ICAM-1)、核因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)及白细胞介素1β(IL-1β)、IL-6和IL-8的含量;荧光探针JC-1检测线粒体膜电位的变化。结果 应用7 mmol/L NAC预处理HUVEC 30 min可明显减轻高糖诱导的HUVEC损伤,使细胞存活率升高,细胞凋亡数量减少,细胞凋亡蛋白cleaved Caspase-3表达减少,胞内活性氧堆积及线粒体膜电位丢失减少(P＜0.01)。NAC能抑制高糖对p-JAK2、p-STAT3表达的上调作用(P＜0.01),同时可抑制高糖诱导的炎症反应,使炎症因子ICAM-1、NF-κB、TNF-α及白细胞介素1β(IL-1β)、IL-6和IL-8的水平下降(P＜0.01)。结论NAC能够通过抑制JAK2/STAT3信号通路对高糖诱导的HUVEC损伤起到保护作用。     

过氧化物酶体增殖物激活受体α的激活可以减轻高糖高脂诱导的心肌细胞凋亡

目的 探讨激活过氧化物酶体增殖物激活受体α(PPARα)对高糖高脂诱导的心肌细胞凋亡的影响.方法 将培养的心肌细胞分为:1)正常组(N组);2)高糖高脂组(H组);3)高糖高脂 PPARα特异性激动剂匹尼尼酸(Wy14643)组(S组).TUNEL法检测细胞凋亡,免疫细胞化学法检测各组细胞PPARα蛋白的表达.结果 H组凋亡细胞较N组增多[(71.34±0.01)% vs (2.50±0.01)%,P＜0.01];H组PPARα蛋白表达下降(0.08±0.02),与N组(0.12±0.02)比较,P＜0.01;PPARα激活剂匹尼尼酸可抑制高糖高脂引起的心肌细胞凋亡,同时伴有培养液中 PPARα蛋白的表达增高(0.16±0.0119 vs 0.08±0.02,P＜0.01).结论 胞核PPARα的表达增高可抑制高糖高脂培养的心肌细胞凋亡,有可能参与机体对高糖高脂血症的自我保护过程.     

自由基清除剂MCI-186对高糖环境中内皮细胞凋亡的干预作用

观察内皮细胞在高糖环境及经肿瘤坏死因子α(TNF-α)诱导后的细胞凋亡情况，并加入自由基清除剂MCI-186予以干扰。检测细胞DNA裂解片段及凋亡信号蛋白(caspase)的表达量，以期探讨MCI-186对高糖环境中血管内皮细胞凋亡的干预作用及凋亡机制。发现高糖环境可增加内皮细胞凋亡率，并上调TNF-α诱导的调亡。MCI-186可抑制高糖诱导的细胞凋亡。但无法干预高糖环境中TNF—a诱导的凋亡。     

盐酸小檗碱对高糖、高脂联合诱导内皮细胞分泌TNF-α的干预作用

目的 观察盐酸小檗碱(黄连素)对高糖、高脂联合诱导下血管内皮细胞分泌肿瘤坏死因子(TNF-α)的干预作用及对内皮细胞的保护作用.方法 体外培养人脐静脉内皮细胞,建立Glu+ ox-LDL损伤HUVECs模型.实验分组:对照组:DMEM培养液+l0％胎牛血清;模型组:Glu40 mmol/L+ ox-LDL 100 mg/L;黄连素低组:Glu 40 mmol/L+ox-LDL 100 mg/L+黄连素1.25 μg/ml;黄连素中组:Glu 40 mmol/L+ ox-LDL 100 mg/L+黄连素2.5 μg/ml;黄连素高组:Glu 40 mmol/L+ ox-LDL 100 mg/L+黄连素5μg/ml.检测细胞培养液中的内皮素(ET-1)、一氧化氮(NO)和TNF-α水平.结果 细胞增殖率:Glu 40 mmol/L+ ox-LDL 100 mg/L(模型组)培养HUVECs 24 h,与对照组比较细胞存活率降低(P＜0.01),说明造模成功,同时应用1.25 ～5.0 μg/ml黄连素治疗,细胞存活率明显提高(P＜0.01).NO:模型组NO水平显著低于对照组(P＜0.01),黄连素处理组NO水平显著高于模型组,以5μg/ml剂量组作用最显著(P＜0.01).ET-1:模型组ET-1比对照组显著增高(P＜0.01),黄连素各治疗组比模型组明显降低(P＜0.01).TNF-α:对照组与模型组间内皮细胞培养液中TNF-α水平差别有统计学意义(P＜0.01),黄连素各治疗组比模型组内皮细胞培养液中TNF-α水平明显降低(P＜0.01),但仍高于对照组(P＜0.01).结论 黄连素通过调节内皮NO/ET-1平衡、抗炎作用,改善Glu、ox-LDL联合诱导的内皮细胞损伤.     

球形脂联素对糖基化终产物诱导血管内皮细胞表达巨噬细胞炎性蛋白-1α的影响

目的 观察球形脂联素(gAdAPN)对糖基化终产物诱导血管内皮细胞表达巨噬细胞炎性蛋白(MIP)-1α的影响.方法 体外培养人脐静脉内皮细胞,以人血清白蛋白作为阴性对照,用含不同浓度梯度(100、200、400 mg/L)糖基化终产物的培养液对内皮细胞体外培养60 min及gAdAPN 100 nmol/L作用30 min后再加入400 mg/L糖基化终产物作用内皮细胞60 min,采用Western印迹法检测内皮细胞MIP-1α蛋白的表达.结果 糖基化终产物组内皮细胞MIP-1α蛋白的表达明显高于对照组(P＜0.05),且呈剂量依赖方式;经灰度值分析,各组内皮细胞内MIP-1α蛋白的表达量分别为对照组的1.12倍、1.47倍及1.98倍(P＜0.05).gAdAPN干预组内皮细胞MIP-1α蛋白的表达降低为对照组的1.64倍(P＜0.05).结论 gAdAPN能抑制糖基化终产物诱导血管内皮细胞表达MIP-1α,可能有助于延缓糖尿病大血管病的发生.     

A single injection of adrenergic agonists enhances pineal melatonin production in Turkish hamsters

Abstract: The purpose of this study was to determine whether the pineal gland of Turkish hamsters (Mesocricetus brandti) responds to adrenergic agonists with an increase in melatonin production, and, if it does, whether the sensitivity of the pineal gland to agonists would differ throughout the dark phase. Adult Turkish hamsters weighing 110–210 g received a subcutaneous injection of isoproterenol (ISO, 1 mg/kg B.W.) or norepinephrine (NE, 1 mg/kg B.W.) at different times of night. Animals exposed to LD 16:8 responded to ISO or NE with increased pineal melatonin content only when injected at dawn, when endogenous melatonin is at basal or near-basal levels. When the 8 hr scotophase was entirely replaced with light, the responsiveness to ISO injections at dawn disappeared. In animals exposed to light from 30 min prior to injection to the time of sacrifice, ISO injections increased pineal melatonin content (P < 0.005, three-way ANOVA), which varied, depending on the specific time of injection (effect of time of night, P < 0.05, three-way ANOVA). These results demonstrate that (1) adrenergic agonists enhance the production of pineal melatonin in Turkish hamsters, (2) this stimulatory effect takes place late, but not early in the 8 hr scotophase, and (3) the adrenergic induction of pineal melatonin production in Turkish hamsters requires priming by darkness during the appropriate circadian phase.     

MUTATION FREQUENCY IN NURSES AND PHARMACISTS WORKING WITH CYTOTOXIC DRUGS

Individuals occupationally exposed to cytotoxic drugs may be at risk owing to the effects of these agents on DNA. As an index of DNA damage, in vivo mutations were measured in lymphocytes from 24 oncology nurses or pharmacists and 24 matched controls. Mutation frequency was significantly increased in exposed individuals and appeared to be related to duration of exposure. However, the overall magnitude of the increase was small and its biological significance remains to be determined.     

Immune effector mechanisms in malaria: An update focusing on human immunity

The past decade has witnessed dramatic decreases in malaria‐associated mortality and morbidity around the world. This progress has largely been due to intensified malaria control measures, implementation of rapid diagnostics and establishing a network to anticipate and mitigate antimalarial drug resistance. However, the ultimate tool for malaria prevention is the development and implementation of an effective vaccine. To date, malaria vaccine efforts have focused on determining which of the thousands of antigens expressed by Plasmodium falciparum are instrumental targets of protective immunity. The antigenic variation and antigenic polymorphisms arising in parasite genes under immune selection present a daunting challenge for target antigen selection and prioritization, and is a given caveat when interpreting immune recall responses or results from monovalent vaccine trials. Other immune evasion strategies executed by the parasite highlight the myriad of ways in which it can become a recurrent infection. This review provides an update on immune effector mechanisms in malaria and focuses on our improved ability to interrogate the complexity of human immune system, accelerated by recent methodological advances. Appreciating how the human immune landscape influences the effectiveness and longevity of antimalarial immunity will help explain which conditions are necessary for immune effector mechanisms to prevail.     

Secondary aortoduodenal fistula

Aorto-duodenal fistulae （ADF） are the most frequent aorto-enteric fistulae （80%）, presenting with upper gastrointestinal bleeding. We report the first case of a man with a secondary aorto-duodenal fistula presenting with a history of persistent occlusive syndrome. A 59-year old man who underwent an aortic-bi-femoral bypass 5 years ago, presented with dyspepsia and biliary vomiting. Computed tomography scan showed in the third duodenal segment the presence of inflammatory tissue with air bubbles between the duodenum and prosthesis, adherent to the duodenum. The patient was submitted to surgery, during which the prosthesis was detached from the duodenum, the intestine failed to close and a gastro-jejunal anastomosis was performed. The post-operative course was simple, secondary ADF was a complication （0.3%-2%） of aortic surgery. Mechanical erosion of the prosthetic material into the bowel was due to the lack of interposed retroperitoneal tissue or the excessive pulsation of redundantly placed grafts or septic procedures. The third or fourth duodenal segment was most frequently involved. Diagnosis of ADF was difficult. Surgical treatment is always recommended by explorative laparotomy. ADF must be suspected whenever a patient with aortic prosthesis has digestive bleeding or unexplained obstructive syndrome. Rarely the clinical picture of ADF is subtle presenting as an obstructive syndrome and in these cases the principal goal is to effectively relieve the mechanical bowel obstruction.     

Married to M. tuberculosis: risk of infection and disease in spouses of smear-positive tuberculosis patients

Objectives To quantify the risk of infection and disease in spouses of tuberculosis patients and the extent to which intervention could reduce the risk in this highly exposed group. Methods We compared HIV prevalence, TB prevalence and incidence and tuberculin skin test (TST) results in spouses of TB patients and community controls. HIV‐positive spouses were offered isoniazid preventive therapy (IPT), and TST was repeated at 6, 12 and 24 months. Results We recruited 148 spouses of smear‐positive patients ascertained prospectively and 3% had active TB. We identified 203 spouses of previously diagnosed smear‐positive patients, 11 had already had TB, and the rate of TB was 2.4 per 100 person years(py) over 2 years (95% CI 1.15–5.09). 116 were found alive and recruited. HIV prevalence was 37% and 39% in the prospective and retrospective spouse groups and 17% in controls. TST was ≥10 mm in 80% of HIV negative and in 57% of HIV‐positive spouses ascertained retrospectively; 74% HIV negative and 62% HIV‐positive spouses ascertained prospectively, and 48% HIV negative and 26% HIV‐positive community controls. Of 54 HIV‐positive spouses, 18 completed 6‐month IPT. At 2 year follow‐up, 87% of surviving spouses had TST ≥10 mm and the rate of TB was 1.1 per 100 py (95% CI 0.34–3.29). Conclusions Spouses are a high‐risk group who should be screened for HIV and active TB. TST prevalence was already high by the time the spouses were approached but further infections were seen to occur. Uptake and adherence to IPT was disappointing, lessening the impact of short‐duration therapy.     

Dénervation rénale pour HTA réfractaire sans anesthésie générale : intérêt d’un protocole MEOPA Morphine. Expérience préliminaire

Renal denervation using the technique of radiofrequency is used only recently for the treatment of resistant hypertension. Normally, it is done under general anesthesia because the ablation point technique is painful. We suggest an alternative to general anesthesia comprising an association of morphin 0.1 mg/kg IV to MEOPA (gas combining oxygen and azot protoxyd) delivered through an oxygen mask. Our series includes 12 consecutive patients treated between October 2011 and June 2013, the first five patients (group 1) have received only an hydroxizin and morphin sedation. Every five have felt the ablation painful, in two cases bearable pain (EVA < 5), in three cases intense (EVA > 5) pain leading to increasing doses of morphin, (total dose of 0.25 mg/kg in two cases, 0.17 mg in one case). For the seven following patients, a protocol including hydroxyzin, morphin and MEOPA given through a mask has been set up. Only one patient has felt a mild pain (EVA 5) leading to an increasing dose of morphin (total dose 0.17 mg/kg). None of the six other patients has felt any pain during the procedure. The average dose of morphin is 0.17 mg/kg in group 1, 0.11 mg/kg in group 2. This is a preliminary study; if confirmed, it will allow a lot of hospitals without on-site possibilities of general anesthesia, to realize such procedures. Conclusion: regarding pain, the procedure of renal ablation was well tolerated for six among seven patients receiving the association MEOPA and IV morphin. In contrast, in the five patients treated only with IV morphin, we observed a less good tolerance to pain and the need to increase the doses of IV morphin.     

Scale‐up of TB and HIV programme collaborative activities in Zambia – a 10‐year review

Objective To review the activities, progress, achievements and challenges of the Zambia Ministry of Health tuberculosis (TB)/HIV collaborative activities over the past decade. Methods Analysis of Zambia Ministry of Health National TB and HIV programme documents and external independent programme review reports pertaining to 2000–2010. Results The number of people testing for HIV increased from 37 557 persons in 2003 to 1 327 995 persons in 2010 nationally. Those receiving anti‐retroviral therapy (ART) increased from 143 in 2003 to 344 304 in 2010. The national HIV prevalence estimates declined from 14.3% in 2001 to 13.5% in 2009. The proportion of TB patients being tested for HIV increased from 22.6% in 2006 to 84% in 2010 and approximately 70% were HIV positive. The proportion of the HIV‐infected TB patients who: (i) started on ART increased from 38% in 2006 to 50% in 2010; (ii) commenced co‐trimoxazole preventive therapy (CPT) increased from 31% in 2006 to 70% in 2010; and (iii) were successfully treated increased to an average of 80% resulting in decline of deaths from 13% in 2006 to 9% in 2010. Conclusions The scale‐up of TB/HIV collaborative programme activities in Zambia has steadily increased over the past decade resulting in increased testing for TB and HIV, and anti‐retroviral (ARV) rollout with improved treatment outcomes among TB patients co‐infected with HIV. Getting service delivery points to adhere to WHO guidelines for collaborative TB/HIV activities remains problematic, especially those meant to reduce the burden of TB in people living with HIV/AIDS (PLWHA).     

C-MET在非小细胞肺癌对表皮生长因子受体酪氨酸激酶抑制剂耐药中的作用研究进展

以表皮生长因子受体(EGFR)为靶点的酪氨酸酶抑制剂(TKI)是近年来非小细胞肺癌(NSCLC)治疗的重大突破.但是随着临床的广泛应用,耐药成为新的难点.新近研究已发现对EGFRTKI的耐药产生主要涉及原癌基因C-MET的扩增突变.C-MET是原癌基因,是蛋白产物肝细胞生长因子/离散离子(HGF/SF)的受体,具有酪氨酸酶活性,C-MET基因扩增激活ErbB3-PI3K信号途径导致NSCLC对EGFR-TKI产生耐药,大量研究证实NSCLC患者对EGFR-TKI耐药约20％归因于C-MET基因扩增.     

Artificial intelligence in pancreaticobiliary endoscopy

Artificial intelligence (AI) applications in health care have exponentially increased in recent years, and a few of these are related to pancreatobiliary disorders. AI‐based methods were applied to extract information, in prognostication, to guide clinical treatment decisions and in pancreatobiliary endoscopy to characterize lesions. AI applications in endoscopy are expected to reduce inter‐operator variability, improve the accuracy of diagnosis, and assist in therapeutic decision‐making in real time. AI‐based literature must however be interpreted with caution given the limited external validation. A multidisciplinary approach combining clinical and imaging or endoscopy data will better utilize AI‐based technologies to further improve patient care.