Mutations in the COPII Vesicle Component Gene SEC24B are Associated with Human Neural Tube Defects

Neural tube defects (NTDs) are severe birth malformations that affect one in 1,000 live births. Recently, mutations in the planar cell polarity (PCP) pathway genes had been implicated in the pathogenesis of NTDs in both the mouse model and in human cohorts. Mouse models indicate that the homozygous disruption of Sec24b, which mediates the ER‐to‐Golgi transportation of the core PCP gene Vangl2 as a component of the COPII vesicle, will result in craniorachischisis. In this study, we found four rare missense heterozygous SEC24B mutations (p.Phe227Ser, p.Phe682Leu, p.Arg1248Gln, and p.Ala1251Gly) in NTDs cases that were absent in all controls. Among them, p.Phe227Ser and p.Phe682Leu affected its protein stability and physical interaction with VANGL2. Three variants (p.Phe227Ser, p.Arg1248Gln, and p.Ala1251Gly) were demonstrated to affect VANGL2 subcellular localization in cultured cells. Further functional analysis in the zebrafish including overexpression and dosage‐dependent rescue study suggested that these four mutations all displayed loss‐of‐function effects compared with wild‐type SEC24B. Our study demonstrated that functional mutations in SEC24B might contribute to the etiology of a subset of human NTDs and further expanded our knowledge of the role of PCP pathway‐related genes in the pathogenesis of human NTDs.     

肝X-受体激动剂马尾藻甾醇的体外抗结核活性研究

目的 评价马尾藻甾醇及其24-差向异构体的抗结核活性。方法 运用半制备HPLC法分到马尾藻甾醇异构体,¹H-NMR鉴定异构体的化学结构;采用倍比稀释法测定化合物对结核分枝杆菌Mycobacterium tuberculosis的最小抑菌浓度(minimal inhibit concentration, MIC)、单独使用及联合用药的杀菌作用;采用PNPA法评价化合物对芳胺N-乙酰基转移酶(arylamine N-acetyltransferases, NAT)的抑制作用,并使用ledock go软件进行分子虚拟对接实验。结果 获得24S-和24R-马尾藻甾醇;马尾藻甾醇、24S异构体和24R异构体对M. tuberculosis的MIC值分别是25 μM、12.5 μM和 >100μM,对NAT蛋白的抑制率分别为45.27%、46.95%和29.20%。马尾藻甾醇和24S异构体还能增强氧氟沙星的杀菌作用。从数值上看,24S异构体活性强于24R异构体,分子对接实验提示24S异构体与NAT蛋白之间的结合比24R异构体更牢固。结论 马尾藻甾醇及其24S异构体在体外对结核分枝杆菌具有杀菌作用,可能与细菌胆固醇代谢有关,尚待深入研究。     

Subclinical Hypothyroidism Is Associated With Adverse Prognosis in Heart Failure Patients

Background

It is widely recognized that overt hyper- as well as hypothyroidism are potential causes of heart failure (HF). Additionally it has been recently reported that subclinical hypothyroidism (sub-hypo) is associated with atherosclerosis, development of HF, and cardiovascular death. We aimed to clarify the effect of sub-hypo on prognosis of HF, and underlying hemodynamics and exercise capacity.

Acute effect of propranolol on splanchnic circulation in normal and portal hypertensive rats

Propranolol decreases portal venous pressure in patients with cirrhosis but no method is available in man to study the effect of this beta-blocker on splanchnic organ blood flow. Because in rats, the microsphere method allows evaluation of regional blood flow, the acute effect of propranolol on both splanchnic and systemic circulations was studied in normal rats and in rats with portal hypertension due to portal vein stenosis. Portal venous pressure significantly decreased during propranolol administration in normal (5.6 +/- 1.0-4.7 +/- 1.1 mm Hg; mean +/- SD) as well as in portal hypertensive rats (11.7 +/- 2.3-10.3 +/- 1.8 mm Hg). Propranolol slightly decreased cardiac output and arterial pressure in all rats. Portal tributary blood flow was significantly reduced by propranolol in normal rats (17.4 +/- 3.0-11.3 +/- 2.2 ml/min) and in portal hypertensive rats (23.7 +/- 5.0-16.6 +/- 3.3 ml/min). Accordingly vascular resistance of the different organs in the portal venous territory increased in these rats receiving propranolol. The percentage of the decrease in portal tributary blood flow was significantly more marked than the percentage of reduction in cardiac output in portal hypertensive rats but, in normal rats, these percentages were parallel. Hepatic arterial blood flow did not change or slightly increased and, consequently, hepatic arterial vascular resistance decreased. These findings further clarify the marked effects of propranolol on splanchnic circulation.     

Targeting eradication of chronic myeloid leukemia using chimeric oncolytic adenovirus to drive IL-24 expression

Chronic myeloid leukemia (CML) is a clonal disorder in which cells of the myeloid lineage undergo massive clonal expansion as well as resistance to conventional chemotherapy. Gene therapy hold a great promise for treatment of malignancies based on the transfer of genetic material to the tissues. In this study, we explore whether chimeric oncolytic adenovirus-mediated transfer of human interleukin-24 (IL-24) gene induce the enhanced antitumor potency. Our results showed that chimeric oncolytic adenovirus carrying hIL-24 (AdCN205-11-IL-24) could produce high levels of hIL-24 in CML cancer cells, as compared with constructed double-regulated oncolytic adenovirus expressing hIL-24 (AdCN205-IL-24). AdCN205-11-IL-24 could specifically induce cytotoxocity to CML cancer cells, but little or no effect on normal cell lines. AdCN205-11-IL-24 exhibited remarkable anti-tumor activities and induce higher antitumor activity to CML cancer cells by inducing apoptosis in vitro. Our study may provides a potent and safe tool for CML gene therapy.     

Severe OSA Leading to Long Pauses in 24-h Holter ECG Reversed with CPAP

Introduction Sleep-related problems like sleep apnea are increasing tremendously mostly owing to the disordered lifestyle the present generation is leading which is added like a topping on the base of obesity and metabolic syndrome. The burden on the society is huge taking into consideration the work-time loss and health-related financial issues arising out of these sleep disorders with obstructive sleep apnea (OSA) leading the way. Early diagnosis can prevent several complications of OSA. Cardiovascular diseases, including various arrhythmias, arising due to OSA, are described previously.Case Presentation Herein, an interesting case of OSA, whose pacemaker installation to rectify the long pause could be avoided by simple correction of his OSA using continuous positive airway pressure, is presented. This 49-year-old male patient was diagnosed with severe OSA by using polysomnography and all his significant sinus pauses (highest one with 7.8 sec) during holter ECG monitoring were found to be occurring at night and correcting his OSA with continuous positive airway pressure (CPAP) treatment reverted all those sinus pauses and the need for any further intervention with pacemaker was discarded.Discussion OSA is caused by either partial or complete obstruction of the upper airway, and there is the simultaneously attenuated upper airway dilator muscle tone while the patient is sleeping. The gold standard test designed for the assessment of OSA is polysomnography, as approved by the American Academy of Sleep Medicine and CPAP has been found to be universally beneficial in treating OSA related complications. Physiologically, the ACC/AHA guidelines recommend pacing only in patients with prolonged asymptomatic pauses occurring during wakefulness. This case report proved the above mentioned claim of CPAP treatment.     

肺癌患者放化疗及手术后心律失常监测

目的 研究肺癌患者放化疗或手术后心律失常及心率变异性的变化,为临床提供参考依据.方法 选择我院150例肺癌放化疗或手术后的患者进行常规心电图和24 h动态心电图监测,捕捉心脏损伤的早期心电图改变及心率变异性的变化.结果 其中125例肺癌患者放化疗或手术后出现心律失常（83.33％）;心率变异性正常者45例（30％）,降低者105例（70％）.但是无论肺癌患者是进行放疗、化疗还是手术治疗,都可引起心律失常的发生,且不同治疗方法下的发生率比较,差异无统计学意义（P＞0.05）.结论 早期发现心脏放化疗或术后损伤的心电图改变,有利于后续治疗方案的及时调整,及时实施干预,可预防肺癌患者放化疗或手术后引起的心脏并发症,以改善患者生活质量.     

Decreased Conversion of 25-hydroxyvitamin D3 to 24,25-dihydroxyvitamin D3 Following Cholecalciferol Therapy in Patients with CKD

Background and objectives

Elevated concentrations of fibroblast growth factor 23 (FGF23) are postulated to promote 25-hydroxyvitamin D (25[OH]D) insufficiency in CKD by stimulating 24-hydroxylation of this metabolite, leading to its subsequent degradation; however, prospective human studies testing this relationship are lacking.

Design, setting, participants, & measurements

An open-label prospective study was conducted from October 2010 through July 2012 to compare the effect of 8 weeks of oral cholecalciferol therapy (50,000 IU twice weekly) on the production of 24,25(OH)₂D₃ in vitamin D–insufficient patients with CKD (n=15) and controls with normal kidney function (n=15). Vitamin D metabolites were comprehensively profiled at baseline and after treatment, along with FGF23 and other mineral metabolism parameters.

Results

Vitamin D₃ and 25(OH)D₃ concentrations increased equivalently in the CKD and control groups following cholecalciferol treatment (median D₃ change, 8.6 ng/ml [interquartile range, 3.9–25.6 ng/ml] for controls versus 12.6 ng/ml [6.9–41.2 ng/ml] for CKD [P=0.15]; 25(OH)D₃ change, 39.2 ng/ml [30.9–47.2 ng/ml] for controls versus 39.9 ng/ml [31.5–44.1 ng/ml] for CKD [P=0.58]). Likewise, the absolute increase in 1α,25(OH)₂D₃ was similar between CKD participants and controls (change, 111.2 pg/ml [64.3–141.6 pg/ml] for controls versus 101.1 pg/ml [74.2–123.1 pg/ml] for CKD; P=0.38). Baseline and post-treatment 24,25(OH)₂D₃ concentrations were lower in the CKD group; moreover, the absolute increase in 24,25(OH)₂D₃ after therapy was markedly smaller in patients with CKD (change, 2.8 ng/ml [2.3–3.5 ng/ml] for controls versus 1.2 ng/ml [0.6–1.9 ng/ml] for patients with CKD; P<0.001). Furthermore, higher baseline FGF23 concentrations were associated with smaller increments in 24,25(OH)₂D₃ for individuals with CKD; this association was negated after adjustment for eGFR by multivariate analysis.

Conclusions

Patients with CKD exhibit an altered ability to increase serum 24,25(OH)₂D₃ after cholecalciferol therapy, suggesting decreased 24-hydroxylase activity in CKD. The observed relationship between baseline FGF23 and increments in 24,25(OH)₂D₃ further refutes the idea that FGF23 directly contributes to 25(OH)D insufficiency in CKD through stimulation of 24-hydroxylase activity.     

IL-24基因重组腺病毒载体转染的树突状细胞促进宫颈癌CaSki细胞凋亡

 目的：探讨白细胞介素24(interleukin-24,IL-24) 基因重组腺病毒载体转染的树突状细胞(dendri-tic cells,DCs)是否在体外诱导细胞毒性T淋巴细胞(cytotoxic T-lymphocytes,CTLs)对宫颈癌CaSki细胞的杀伤效应。方法：分离培养小鼠未成熟DCs,将已成功构建的带有IL-24基因的重组腺病毒感染体外培养的小鼠未成熟DCs,提取CaSki细胞裂解物负载DCs,制备DC疫苗,Western blotting检测IL-24蛋白的表达;PE-Annexin V和Western blotting检测细胞凋亡及cleaved caspase-3表达;克隆形成实验检测克隆形成能力;裸鼠荷瘤模型体内研究带有IL-24基因的重组腺病毒载体转染DCs在体外诱导的CTLs对宫颈癌CaSki细胞的杀伤效应。结果：Western blotting检测IL-24蛋白高表达;DCs疫苗诱导产生的CTLs后与CaSki细胞共培养,对CaSki细胞生长具有明显的抑制作用, DCs疫苗诱导产生的CTLs促进CaSki细胞凋亡,增加cleaved caspased-3蛋白表达,克隆形成实验检测该疫苗具有抑制CaSki细胞形成克隆的能力。裸鼠的成瘤实验表明,带有IL-24基因的重组腺病毒载体转染DCs可以抑制体内肿瘤的形成,促进肿瘤细胞凋亡。结论：IL-24基因重组腺病毒感染DCs制备的DCs疫苗可诱导CTLs,从而抑制宫颈癌CaSki细胞增殖,并促进其凋亡。