miR-200b调控肌球蛋白轻链激酶/磷酸化肌球蛋白轻链信号通路对肠上皮紧密连接的影响

目的探究mi R-200b对肠上皮紧密连接的影响及作用机制。方法利用阴性对照慢病毒及表达mi R-200b的慢病毒感染Caco-2细胞形成NC株和200b株,以10 ng/m L肿瘤坏死因子(TNF-α)处理建立体外肠上皮损伤模型,并分为NC组、NC+TNF-α组、200b组和200b+TNF-α组。测量4组细胞对肠上皮跨膜电阻抗(TEER)及对异硫氰酸荧光素-右旋糖酐(FITC-dextran)的通透性;Western blot检测肌球蛋白轻链激酶(MLCK)及磷酸化肌球蛋白轻链(P-MLC)表达。结果与NC组相比,NC+TNF-α组TEER值降低、对FITC-dextran通透量显著增高、MLCK和P-MLC表达上调,差异均有统计学意义(P?0.05)。与NC+TNF-α组相比,200b+TNF-α组TEER显著升高、FITC-dextran通透量显著降低、MLCK和P-MLC表达显著下调,差异均有统计学意义(P?0.05)。结论 mi R-200b可调控MLCK/P-MLC信号通路修复TNF-α导致的肠上皮紧密连接损伤。     

脑脊液游离轻链检测在儿童中枢神经系统感染中的应用

目的探讨脑脊液(CSF)游离轻链检测在儿童中枢神经系统感染中的应用价值。方法用BN-Ⅱ特定蛋白分析仪检测24例病毒性脑炎患儿、7例化脓性脑膜炎患儿及12例CSF正常患儿的血清及CSF中白蛋白(ALB)、免疫球蛋白G(IgG)、游离轻链;用免疫固定电泳检测IgG寡克隆区带;计算白蛋白商值(QALB)、IgG商值(QIgG)、Kappa商值(Qκ)、Lamda商值(Qλ),并进行对比分析。结果所有患儿CSF寡克隆区带均为阴性;病毒性脑炎组和化脓性脑膜炎组患儿的QALB、QIgG、Qκ、Qλ显著高于CSF正常对照组(P均<0.05),化脓性脑膜炎组QALB、QIgG显著高于病毒性脑炎组(P均<0.01),病毒性脑炎组Qκ、Qλ与化脓性脑炎组比较,差异均无统计学意义(P>0.05);QIgG、Qλ随血脑屏障损害程度的加深呈递增趋势(P均<0.05);所有研究对象的QALB与QIgG、QALB与Qκ、QALB与Qλ、QIgG与Qκ、QIgG与Qλ、Qκ与Qλ均有相关性(r=0.320～0.814,P均<0.05)。结论游离轻链检测对儿童化脓性脑膜炎和病毒性脑炎的诊断有较大帮助,且Qλ能较好的反映患儿血脑屏障的损伤程度。     

脑源性神经生长因子经自噬对胚鼠神经元缺氧损伤的保护作用

目的 探讨脑源性神经生长因子(BDNF)对胚鼠缺氧神经元是否有保护作用及其机制是否与激活自噬有关.方法 将SD大鼠胚鼠(孕17 ～ 19 d)的大脑皮质神经元在体外进行原代细胞培养,并进行神经元鉴定.培养7～10d,选取生长良好的神经元用于前后两部分实验.1.第一部分随机分为3组,对照组:不加入药物,只作缺氧处理;3-甲基腺嘌呤组(3-MA组):提前加入不同浓度的3-MA后作缺氧处理,依次为5 mmol·L-1 3-MA组、10 mmol·L-1 3-MA组、20 mmol·L-1 3-MA组;BDNF组:提前加入不同浓度的BDNF后作缺氧处理,依次为50μg· L-1 BDNF组、100 μg·L-1 BDNF组、200 μg·L-1 BDNF组.观察不同剂量3- MA、BDNF对缺氧神经元损伤的作用.之后以细胞计数盒-8(CCK-8)测定各组细胞活力,确定干预缺氧神经元的最佳药物浓度.2.第二部分分为3组,对照组:单纯缺氧,不加药物;100 μg·L-1 BDNF组:加入100 μg·L-1BDNF;10 mmol·L-1 3-MA组:加入10 mmol·L-1 3-MA.免疫蛋白印迹法检测3组缺氧神经元在不同缺氧时间点细胞自噬微管相关蛋白轻链3(LC3)的表达情况,以LC3Ⅱ/actin的相对表达量判断自噬发生的程度.结果 1.免疫荧光鉴定:与未缺氧神经元比较,缺氧神经元突触回缩,细胞网状结构被破坏.2.神经元细胞活力:50 μg·L-1BDNF组缺氧神经元细胞活力最强,100 μg·L-1BDNF组缺氧神经元细胞活力其次(Pa＜0.05);随3-MA剂量加大,各剂量组神经元活力明显降低.3.免疫印迹:于缺氧1h、3h、5h,100 μg·L-1 BDNF组缺氧神经元的LC3表达量,均较对照组相应时间点明显上调(Pa＜0.05).结论 BDNF通过自噬途径对缺氧损伤神经元起保护作用.     

Relationship between the degree of injury at operation and the change in antimyosin antibody titer in the postpericardiotomy syndrome

Summary Successive measurements of cardiac myosin light chain I (MLC), creatine kinase isoenzyme MB (CKMB), and the titer of antimyosin antibody (AMA) were performed prospectively in 19 patients following open heart surgery. Seven of these patients showed the postpericardiotomy syndrome (PPS). No differences in serum concentrations of MLC or CKMB were observed between the patients with and without PPS, and all patients in both groups had abnormal MLC values after surgery. However, only patients with PPS had significantly elevated AMA titers. The maximum AMA titer was significantly correlated with the severity of the effusion. These data suggest that PPS is unrelated to the severity of myocardial injury during operation. Furthermore, the AMA titer may be useful as one of the indicators for determining the patient's clinical condition.     

β抑制蛋白2及微管相关蛋白轻链3在大鼠肾脏缺血再灌注损伤中的改变及意义

目的 探讨β抑制蛋白2(β-arrestin2)及微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3)在急性肾脏缺血再灌注损伤中的表达及与肾脏损害程度的相关性.方法 选用生后3~4周的雄性SD大鼠,随机分为正常组、假手术组、急性缺血再灌注损伤组.通过右侧肾脏切除,无创动脉夹夹闭左侧肾动脉45 min之后松开动脉夹,恢复肾脏血流,建立肾脏急性缺血再灌注损伤模型.并在恢复肾脏血流后12、24、36、48、72、96 h取肾脏及血液样本.采用免疫组织化学方法及West-ern blot方法检测各组肾组织中β-arrestin2及LC3蛋白的表达水平,检测各组的肾功能,并对各组肾脏病理学进行评分.结果 与正常组及假手术组相比,缺血再灌注损伤组血肌酐及肾脏病理学评分均有显著升高,其中肾脏损伤程度以缺血再灌注损伤后24 h最为明显;β-arrestin2及LC3蛋白在正常组及假手术组肾脏中的表达较少,在缺血再灌注损伤后的肾脏中表达显著升高,其中以缺血再灌注损伤后12 h时表达上调最为显著;β-arrestin2及LC3的表达改变先于肾脏病理改变,并且与肾脏损害程度呈正相关(r=0.821,P<0.05;r=0.913,P<0.05).结论 在肾脏急性缺血再灌注损伤时,β-arrestin2可能作为一个上游调控蛋白,通过对自噬的调节参与急性肾损伤的病理过程.     

Change of bilirubin photoisomers in the urine and serum before and after phototherapy compared with light source

BACKGROUND: The clinical effect of phototherapy for neonatal hyperbilirubinemia is based on the production and elimination of cyclobilirubin. Generally, the clinical effect of light sources is estimated by the reduction in the total serum bilirubin level. One procedure with less invasiveness than blood collecting is urine collection. Whether the effectiveness of light sources used for phototherapy could be assessed using measurements of bilirubin photoisomers in urine was studied. METHODS: This study was a retrospective analysis of 38 term infants with hyperbilirubinemia who underwent phototherapy. Bilirubin fractions in serum and urine before and 24 h after the phototherapy were measured by high-performance liquid chromatography. The light sources used for the phototherapy were blue-white light (n = 11), Biliblanket plus high output (n = 13) or green light (n = 14). The relationships between serum and urine bilirubin photoisomers after phototherapy and whether the levels of urine bilirubin photoisomer are affected by the light sources with different wavelength characteristic were analyzed. RESULTS: There was no correlation between serum (ZE)-bilirubin and urine configurational isomers, but a weak positive correlation between serum (EZ)-cyclobilirubin and urine structural isomers after phototherapy. Although serum (ZE)-bilirubin levels depended on the wavelength characteristic of each light source during phototherapy, the urine configurational isomer levels did not depend on it. The increase in serum (EZ)-cyclobilirubin levels and the urine structural isomer levels were mostly in agreement. CONCLUSIONS: The urine bilirubin structural isomers may be used to estimate the serum (EZ)-cyclobilirubin levels and to evaluate the clinical effects of light sources.     

Therapeutic effect of turquoise versus blue light with equal irradiance in preterm infants with jaundice

AIM: To compare the efficiency of turquoise light with that of TL52 blue in treatment of preterm infants with jaundice at the same level of body irradiance. METHODS: Infants with gestational age 28-37 weeks and non-haemolytic hyperbilirubinemia were treated for 24 h with either turquoise light (OSRAM L18W/860 fluorescent lamps) or blue light (Philips TL20W/52 fluorescent lamps). The concentrations of serum total bilirubin and bilirubin isomers were measured by the Vitros routine method and by HPLC, respectively. RESULTS: The decrease in serum concentrations of total bilirubin, total bilirubin isomers and the toxic Z,Z-bilirubin was greatest for infants treated with turquoise light. Further, the increase in Z,E-bilirubin was smaller and there was a trend towards a higher rise in E,Z-bilirubin. CONCLUSIONS: Turquoise light has a greater bilirubin reducing effect than TL52 blue light with equal irradiance, expressed both by serum total bilirubin, total bilirubin isomers and Z,Z-bilirubin, i.e. the turquoise spectral range is more efficient than the blue. This is in accordance with deeper penetration into the skin, lower production of the Z,E-bilirubin and greater production of E,Z-bilirubin and lumirubin, in infants under turquoise light. This suggests, given equal irradiances, that light in the turquoise spectral range is preferable to the TL52 blue in treatment of newborn jaundiced infants.     

Effect of light reduction on the incidence of retinopathy of prematurity

The effect of ocular protection on the incidence of retinopathy of prematurity was tested in 188 newborns weighing less than 1600 g in a randomised controlled trial. No effect of ambient light reduction on the incidence of retinopathy of prematurity was shown.     

极长链酰基辅酶A脱氢酶缺乏症研究进展

极长链酰基辅酶A脱氢酶缺乏症是一种较罕见的脂肪酸代谢障碍疾病,根据起病年龄和临床表现分为三型：心肌病型、肝型、肌病型。心肌病型病情重,病死率高。临床诊断可通过血串联质谱(MS/MS)检测血肉豆蔻烯酰基肉碱(C14:1)水平进行,进一步确诊可通过基因诊断、酶学分析及脂肪酸氧化流量分析。治疗上主要包括避免空腹,减少长链脂肪酸的摄入,补充中链甘油三酯等。     

自噬与全身型幼年特发性关节炎发生的关系及其相关机制的初步研究

目的 通过研究全身型幼年特发性关节炎（sJIA）患儿外周血淋巴细胞中微管相关蛋白轻链3-Ⅱ（LC3-Ⅱ）、髓样细胞分化因子88（MyD88）及T细胞受体信号抑制因子1（STS-1）的表达,探讨自噬在sJIA发生发展中的作用。方法 26例sJIA患儿及26例健康体检的儿童（对照组）纳入研究。采用Western blot检测外周血淋巴细胞中LC3-Ⅱ、STS-1及MyD88蛋白的表达水平,免疫荧光法检测LC3-Ⅱ在淋巴细胞胞浆中的表达,并采用Pearson相关分析法进行各指标间的相关性分析。结果 sJIA组LC3-Ⅱ、STS-1、MyD88表达均较对照组显著增高,差异有统计学意义（P < 0.05）。sJIA组LC3-Ⅱ表达与MyD88表达呈正相关（r=0.478,P < 0.05）;STS-1表达与MyD88表达亦呈正相关（r=0.817,P < 0.01）。结论 sJIA患儿外周血淋巴细胞LC3-Ⅱ高表达,提示sJIA的发生发展与自噬过度表达有关;STS-1可能通过激活某些信号通路诱导自噬的发生;MyD88可能通过Toll样受体信号通路参与自噬的发生。     

Iatrogenic radiant heat burns in severely asphyxic newborns

We report two cases of newborns who developed second-degree burns following resuscitation under infra-red heating lamps. Both infants were asphyxic and suffered from insufficient peripheral circulation which, combined with the long duration of the exposure to the light, contributed to the development of the lesions. Both infants died shortly after birth for reasons other than the burns.     

Empyema thoracis

Parapneumonic effusion and empyema thoracis remains a significant source of morbidity in children, though the overall incidence of empyema thoracis has decreased in the past two decades. These conditions pose a dilemma regarding evaluation and treatment for the treating physician. This article discusses the practical strategies in the management of empyema thoracis in children.     

用聚合酶链反应研究先天性畸形与巨细胞病毒感染的关系

应用聚合酶链反应（ＰＣＲ）方法检测畸形和非畸形新生儿死亡病例尸检的石蜡包埋病理组织标本中的巨细胞病毒核糖核酸（ＣＭＶ－ＤＮＡ）片断，研究先天性畸形与宫内ＣＭＶ感染的关系以及胎儿各脏器对ＣＭＶ的易感性。结果显示：（１）畸形组中ＣＭＶ的阳性率高达４２．１９％（２７／６４），明显高于非畸形组的４．６９％（３／６４），差异有非常显著意义（Ｘ￣２＝２５．０７８，Ｐ＜０．００１），提示宫内ＣＭＶ感染与胎儿畸形密切相关；（２）神经系统畸形病例ＣＭＶ的阳性率与循环系统畸形组比较，差异有显著意义（Ｘ￣２＝６．６２６，Ｐ＜０．０５）。提示ＣＭＶ感染可能导致小头畸形、脑积水和脊柱裂等畸形；（３）脑和肝脏是最易受ＣＭＶ感染的脏器；（４）肝胆异常与ＣＭＶ感染有关。本研究说明，先天性ＣＭＶ感染易导致胎儿神经系统崎形及肝脏损害。     

Light reduction and the electroretinogram of preterm infants

AIMS—To examine the effects of light on retinal development and function in preterm infants as measured by the electroretinogram (ERG). Secondary outcomes included visual acuity testing, the incidence of retinopathy of prematurity, and general wellbeing, reflected in feeding tolerance, rate of weight gain, and length of hospital stay.METHODS—Eligibility criteria for enrolment were birthweight ? 1250 g and gestational age ? 31 weeks. Sixty one infants were randomly allocated by 6 hours after birth to a control or treatment group which wore 97% light filtering goggles for a minimum of four weeks or until the infant reached 31 weeks postmenstrual age.RESULTS—There were no significant differences between the two groups in the numbers of electroretinograms performed at 36 weeks of postmenstrual age. Although the sample size was not large enough to exclude clinically important differences in secondary outcomes, no significant differences were observed between the groups in visual acuity testing at 4-6 months corrected age, incidence of retinopathy of prematurity, weight gain, or length of stay.CONCLUSION—These data support the safety and feasibility of this intervention. A much larger study will be needed to determine whether light reduction to the eyes of very low birthweight infants will reduce the incidence of retinopathy of prematurity or enhance general wellbeing.     

Nested amplification protocol for the detection of Mycobacterium tuberculosis

Several methods for rapid diagnosis of tuberculosis have been devised through DNA amplification. However, the chemically strong cell wall of the species, the presumptively low numbers of organisms and their uneven distribution in clinical samples, and the lack of a "gold standard" for diagnosing tuberculosis, have hindered the routine clinical use of this method. In a pediatric patient group, these factors are more perplexing. To circumvent these problems, we made use of nested amplification and developed a standard protocol for extracting DNA from various forms of clinical samples which were suitable to our clinical laboratory. It is our impression that the overall sensitivity, including technical bias accompanying this method, is equal to, or at least greater than, that of culture. Most notably, the rapidity in obtaining results and the simplicity in handling, storage and transfer of samples are the principal advantages of this method.     

蓝光照射治疗新生儿黄疸的护理体会

目的采用蓝光照射治疗新生儿黄疸，精心护理使疗效提高，避免发生副作用。方法对198例新生儿高胆红素血症患儿采用蓝光治疗，进行细心观察与护理。结果198例新生儿黄疸经蓝光治疗均痊愈出院，无不良后遗症。结论蓝光照射能有效降低新生儿黄疸患儿的血清胆红素，而正确、精心的护理则是蓝光治疗的关键。     

不同光源对新生儿高胆红素血症的作用评价

目的　 评价不同光源的作用。 方法 　生后 49～ 72h ,血清总胆红素 >2 0 5 μmol/L之足月高胆儿 ,随机分为蓝光组、蓝绿光组及绿光组各 2 0例 ,持续光疗 48h。光强度 (单位 μw·cm2 ·nm)蓝光 12～ 16,蓝绿光 8～ 12 ,绿光 5～ 8。光疗前后分别检血细胞、血生化及内分泌激素。 结果 　与光疗前比较 ,光疗后蓝光及蓝绿光组血红细胞减少 ;三组血胆红素均显著下降 ,以蓝光组为著 ,蓝绿光组次之 ,绿光组不像文献所报导有效 ;三组对其他血细胞、血生化及内分泌激素均无影响。 结论 　除蓝光可致轻度贫血外 ,无其他副作用。但比较不同光源对降低血胆红素的疗效 ,必须要光谱纯度、光强度、半衰期等条件较一致时 ,才能作出科学的评价。     

Familial neonatal SIDS revealing carnitine-acylcarnitine translocase deficiency

A patient with a severe phenotype of carnitine-acylcarnitine translocase deficiency (CATR)(McKusick 212138) is reported. Prior to birth, a defect in β-oxidation was suspected because of neonatal death of six siblings. Dietary treatment during neonatal adaptation and the subsequent six months of life and a trial of carnitine supplementation are reported. The rapidity with which long chain fatty acid metabolites can accumulate and induce secondary carnitine deficiency within a few hours after birth in an infant with CATR is noteworthy. Conclusion High rates of glucose suppressed neonatal lipolysis in this infant, but did not seem sufficient to avoid secondary carnitine deficiency as in severe forms of CATR. Therefore simultaneous use of insulin and glucose may be necessary to control neonatal lipolysis. Carnitine supplementation and the possible adverse effects of MCT systematically administrated, should be further assessed in patients with CATR. Received: 13 April 1999 / Accepted: 26 August 1999