Identification of two mutations in a compound heterozygous child with dihydrolipoamide dehydrogenase deficiency

An infant girl with elevated blood lactate, pyruvate, and plasma branched-chain amino acids was diagnosed with dihydrolipoamide dehydrogenase (E3; dihydrolipoamide: NAD+ oxidoreductase, EC 1.8.1.4) deficiency. Activities of the pyruvate dehydrogenase complex and E3 from patient were 26 and 2% of controls in blood lymphocytes, and 11 and 14% in cultured skin fibroblasts, respectively. Western blot analysis demonstrated that the amount of E3 protein in fibroblasts from the patient and her father was about half of controls, while Northern blot analysis showed normal amounts of E3 RNA. DNA sequencing of cloned full-length E3 cDNAs from the patient revealed two mutations in separate alleles. One is a single base insertion of an extra adenine in the last codon of the leader peptide sequence (TAC-->TAAC) leading to a nonsense mutation which results in the premature termination of the precursor E3 polypeptide (Y35X). The other is a missense mutation due to substitution of guanine for adenine, causing an Arg-->Gly substitution at amino acid 460 of the mature protein (R460G) which triggers the loss of E3 activity probably by structural change in the E3 dimer. DNA sequencing of E3 cDNAs from the parents demonstrated that the nonsense mutation was inherited from the father and the missense mutation was inherited from the mother.      

Membrane ionic currents and properties of freshly dissociated rat brainstem neurons

It is well known that neuronal firing properties are determined by synaptic inputs and inherent membrane functions such as specific ionic currents. To characterize the ionic currents of brainstem cardio-respiratory neurons, cells from the hypoglossal (XII) nucleus and the dorsal motor nucleus of the vagus (DMX) were freshly dissociated and membrane ionic currents were studied under whole-cell voltage and current clamp. Both of these neurons showed a TTX-sensitive Na⁺ current with a much larger current density in XII than DMX neurons. This Na⁺ current had two (fast and slow) distinct inactivation decay components. The ratio of the magnitudes of the fast to slow component was roughly two-fold greater in DMX than in XII cells. Both DMX and XII neurons also showed a high voltage-activated Ca²⁺ current, but this current density was significantly greater (three-fold) in DMX than XII neurons. A relatively small amount of low-voltage activated Ca²⁺ current was also observed in DMX neurons, but not in the majority of XII cells. A transient and a sustained outward current components were observed in DMX cells, but only sustained currents were present in XII neurons. These outward currents had a reversal potential of about − 70 mV with 3 mM external K⁺ and −30 mV with 25 mM K⁺, and substitution of K⁺ with cesium and tetraethylammonium suppressed more than 90% the outward currents, indicating that most outward currents were carried by K⁺. The transient outward current consisted of two components with onesensitive to 4-aminopyridine and the other to intracellular Ca²⁺. In XII neurons, BRL 38227 (lemakalim), an ATP-sensitive K⁺ (K_ATP) channel activator, increased the sustained K⁺ currents by 10% of control, and glibenclamide, a K_ATP channel blocker, decreased the sustained K⁺ currents by 20%. Evidence for the presence of an inward rectifier K⁺ current was also obtained from both XII and DMX neurons. These results on XII and DMX neurons indicate that (1) the methods used to dissociate neurons provide a useful means to overcome voltage clamp technical difficulties; (2) ion channel characteristics such as density and biophysical properties of DMX neurons are very different from those of XII neurons; and (3) several newly discovered membrane ionic currents are present in these cells.     

Brain abscess. A study of 45 consecutive cases

Clinical features, findings of diagnostic studies, results of therapy, and prognostic factors were analyzed in 45 patients with brain abscesses. The number of patients diagnosed yearly has increased since CT scanning became available, but despite the enhanced sensitivity, the time from either onset of symptoms or hospital admission until initiation of therapy was not decreased and there was no dramatic effect upon morbidity or mortality in this series. Infections of paranasal sinuses, ears, lungs, and odontogenic foci were predisposing factors in approximately 70% of cases. Single abscesses, present in 75% of patients, were distributed equally in both hemispheres, with more than half in the frontal and parietal lobes. Common signs and symptoms included headache, fever, chills, seizures, nausea, vomiting, altered sensorium, nuchal rigidity, and localizing neurologic signs. Blood cultures were positive in 11%. Lumbar puncture rarely provided data from which a diagnosis could be established; CSF cultures were positive in only 7% of patients, and there was a 15% temporally associated incidence of brain herniation and death. Diagnostic information was most readily obtained using imaging techniques such as CT and 99mTc scanning, and arteriography was invasive and of no added value. CT scans are however, often initially negative in patients presenting with clinical signs of meningitis presumably following rupture of an abscess into the subarachnoid space, and the average time for changes to appear on CT scan is 9 days. It is, therefore, recommended that when the clinical assessment suggests the possibility of brain abscess the patient be treated empirically with antibiotics and that lumbar puncture be performed only after thoughtful assessment of the risk-to-benefit ratio for each patient. Causative organisms were isolated from more than 80% of abscesses despite prior antibiotic treatment; more than half grew a single pathogen, most commonly streptococci. Anaerobic and microaerophilic bacteria accounted for 62% of all isolates, and were the only organisms in 33% of patients. Computerized tomographic scans in 30 patients showed "ring-enhancing" lesions, nodular enhancement, or areas of low attenuation. Complete resolution of abscesses on CT scans rarely occurred during hospitalization and took as long as 5 months. Decrease in the size of abscesses on CT scan correlated well with clinical improvement and was seen within a week when abscesses were excised, but was often not obvious for 6 to 8 weeks if antibiotics were used alone.(ABSTRACT TRUNCATED AT 400 WORDS)     

两种血浆代用品对凝血功能影响的比较

目的：比较血定安与血代两类药物对患凝血功能的影响。方法：选择60例ASA（美国麻醉医师学会）1－2级手术患随机分为两组（血定安组与血代组）。入室后采集静脉血测定凝血酶原时间、纤维蛋白酶定量、血小板计数、血球压积和血钙含量。分别输入血定安与血代1000ml后,再次于非输液侧采集静脉血,监测上述指标。结果：比较两类药物输液前后血球压积P＜0．05,差异显性。其余各项指标均在正常范围内变化。结论：输注血浆代用品后对凝血机制无明显,仅对血液有稀释作用。通过稀释血液,还能提高血液的运氧能力。     

肾综合症出血热血清蛋白与肝肾功能及临床关系的研究

目的 了解肾综合症出血热（HFRS）各期血清蛋白的变化特点及其与肝、肾功能的关系。方法 采用瑞士产COBAS MIRA PLUS CC全血动生化分析仪及日本产MA－4210尿液分析仪分别检测同时的空腹静脉血及晨尿。结果 150例HFRS患者217次检测血清总蛋白（T）平均为63.51g/L,白蛋白（A）为36.29g/L,球蛋白（G）为27.22g/L,A/G=1.33。其中A／G为1.49-1.0者137例（63.3%）,A／G＜1.0者27例（12.44%）,A／G≥1.5者53例（24.42%）。结论 HFRS患者有75.58%存在着低白蛋白血症,并贯穿于各期,异常程度与病情轻重呈正相关,主要原因是肾脏损害,大量白蛋白从尿中丢失,与肝脏的合成功能关系不大。     

葛根素对降低2型糖尿病患者微量白蛋白尿的临床观察

目的：观察葛根素对2型糖尿病早期肾脏病变的疗效。方法：对73例2型糖尿病24h尿微量白蛋白（UAE）在30－300mg/24h者随机分成两组,治疗组37例,每天静滴葛根素500mg,共2周;对照组用生理盐水静滴,共2周。观察血糖及尿UAE变化。结果：两组治疗前后比较差异有显著性意义,P＜0．01。结论：葛根素对2型糖尿病早期肾脏病变疗效明显,且无明显毒副反应。     

一氧化氮对犬下食管括约肌压力的作用

目的 :应用苄基二甲基十四烷氯化铵 (benzyldim ethyltetradecyl amm onium chloride,BAC)建立犬下食管括约肌(L ES)无神经动物模型 ,研究一氧化氮 (NO)对 L ES压力的作用。方法 :将 BAC环周注入犬 L ES,对照组注入等量生理盐水 ,均于注射前及注射后 6周测定 L ES压力 ;并观察 L -精氨酸、D -精氨酸、硝普钠及一氧化氮合酶 (NOS)抑制剂 N-硝基 - L -精氨酸 (L - NNA)对 L ES压力的影响 ;此外还测定了两组犬 L ES中 NO含量和 NOS活性。结果 :BAC处理组 L ES压力 [(4 2 .43±4.19) m m Hg,1m m Hg=0 .133 k Pa]显著高于对照组 [(2 2 .71± 5 .19) mm Hg]。 L -精氨酸可使对照组 L ES压力降低 ;L - NNA使其增高 ,但对 BAC处理组 L ES压力均无影响。硝普钠可降低两组犬 L ES压力。对照组 L ES中 NO为 (6 .0 5 8± 2 .0 6 7)μm ol/g,NOS为 (1.45 8± 0 .146 ) U /mg;而 BAC处理组 NO为 (1.797± 0 .873)μmol/g,NOS为 (0 .46 3± 0 .0 39) U /m g,均较对照组显著降低 (P<0 .0 1)。 结论 :BAC可使犬 L ES压力增高 ,其机制可能与 L ES局部 NO减少有关。     

Updates on bacterial resistance and empirical antibiotics treatment of complicated acute appendicitis in children

Objective: Through historical comparison with our previous study published 10 years ago, this paper aims to provide latest analysis of local bacteriology of acute complicated appendicitis and evaluate the effects of early escalation of potent antibiotics on course of postoperative recovery.Methods: A 5-year retrospective review of all children receiving emergency laparoscopic appendicectomies for acute appendicitis from December 2014 to November 2019 was conducted.Results: 257 cases of acute appendicitis were included, 126 were complicated appendicitis (38 gangrenous, 88 ruptured). 96 had positive peritoneal swab culture, 53 (42.1%) grew resistant bacterial strains, including extended spectrum beta-lactamase producing E. coli (ESBL E. coli), Pseudomonas aeruginosa, against traditional empirical triple antibiotics. The prevalence had significantly increased over the past decade (p = 0.008). In our patients, piperacillin/tazobactam, ertapenem, gentamicin provided coverage of 69.8%, 45.3% and 45.3% respectively. For patients with early escalation of postoperative antibiotics, no statistical significance was identified in terms of postoperative complications (p = 0.883), or duration of antibiotics (p = 0.0615).Conclusion: Growing prevalence of resistant strains were observed over the decade. Piperacillin/tazobactam provided the best coverage (69.8%) against resistant bacterial strains in our patients. Early escalation of antibiotics failed to reduce postoperative complications and antibiotics duration.Type of Study: Clinical Research, Retrospective Historical Comparative StudyLevel of Evidence: Level III     

阴道炎患者HPV6/11-DNA定量分析

目的：了解女性阴道炎患者人乳头瘤病毒6／11型（HPV6／11）的感染情况。方法：荧光定量聚合酶链反应（FQ－PCR）技术检测258例门诊患者阴道分泌物中HPV6／11的病毒拷贝数。结果：共检出HPV6／11阳性者94例,阳性率36．43％,拷贝数在10^5以上者78例,占阳性者中82．98％,40岁以上年龄组阳性率高且病毒复制量均在10^5以上。结论：（1）中老年阴道炎患者就诊晚,感染重。（2）FQ－PCR检测HPV敏感,快速,准确,特别是其定量特点对临床很有意义。     

淋病奈瑟氏菌中国流行株gyrA和parC基因与喹诺酮类耐药性的相关性

目的　调查中国地区淋病奈瑟氏菌 (NG)对喹诺酮类的耐药性状况 ,探讨高水平耐药的基因突变株对喹诺酮类耐药机制的作用 .方法　对 9a来临床分离保存淋球菌流行株进行了体外环丙沙星 (CIP)药敏实验 .筛选出 76株高水平CIP耐药株 ,通过 PCR扩增了其中的 1 8株 NG的 gyr A和par C喹诺酮耐药决定区基因 ,并对扩增子直接测序 .通过N he I酶切、脉冲电场凝胶电泳 (PFEG)分析了这些菌株的遗传关系 .结果　环丙沙星的耐药检出率有逐年增高的趋势 ,MIC≥ 1 .0 mg· L- 1的菌株由 1 993年的 5 .1 %上升到 2 0 0 1年的 2 1 .4% .对照组中的 1 8株敏感菌只有两株发生了 gyr A的单一突变 ,未发现 par C变异 ;而耐药组中 89% (1 6 )菌株的gyr A发生了 Ser- 91向 Phe的单一突变株或 (和 ) par C的双突变 .88.9% (1 5 )的菌株的 gyr A发生了 Ser- 91向 Phe和 Asp-95向 Gly的突变 .在发生 par C突变中 72 %属于 Asp- 86向Asn突变 .PFEG分析发现 1 0株具有完全一致的 gyr A和par C突变模式 .同时也发现来自不同地区的菌株的 PFEG指纹图是不同的 .结论　研究表明 ,在中国 NG流行株对喹诺酮类耐药率呈增长趋势 ;NG流行株对喹诺酮产生耐药性与gyr A和 par C基因双重突变有着密切的关系