Recombinant adeno-associated virus (rAAV) is an attractive tool for basic science and translational medicine including gene therapy, due to the versatility in its cell and organ transduction. Previous work indicates that rAAV transduction patterns are highly dependent on route of administration. Based on this relationship, we hypothesized that intraperitoneal (IP) administration of rAAV produces unique patterns of tissue tropism. To test this hypothesis, we investigated the transduction efficiency of 12 rAAV serotypes carrying an enhanced green fluorescent protein (EGFP) reporter gene in a panel of 12 organs after IP injection. Our data suggest that IP administration emphasizes transduction patterns that are different from previously reported intravascular delivery methods. Using this approach, rAAV efficiently transduces the liver, pancreas, skeletal muscle, heart and diaphragm without causing significant histopathological changes. Of note, rAAVrh.10 showed excellent muscle transduction following IP administration, highlighting its potential as a new muscle-targeting vector.
Background and purposeCurrently, there are limited options for treatment of postherpetic neuralgia (PHN) patients in China. While pregabalin is an effective treatment option for PHN in several countries, there is limited information on its efficacy in Chinese patients. MethodsThis was an 8-week, double-blind, placebo-controlled trial in Chinese patients with PHN randomized (1:1) to pregabalin 300 mg/day or placebo. Primary efficacy endpoint was change from baseline in mean pain score (Daily Pain Rating Scale; 0 = no pain' to 10 = worst possible pain'). Secondary efficacy endpoints included change from baseline in overall pain intensity score, by visual analog scale (VAS; 0 = no pain' to 100 = worst possible pain') and daily sleep interference score (0 = pain does not interfere with sleep' to 10 = completely interferes'). ResultsA total of 220 patients were randomized and received treatment (111 pregabalin and 109 placebo). Improvement in mean pain score with pregabalin was significantly greater than placebo, least squares mean difference (95% CI), -0.71 (-1.08, -0.34); P = 0.0002. Improvements in VAS and sleep interference score at endpoint were significantly greater with pregabalin than placebo, least squares mean difference (95% CI), -8.18 (-11.99, -4.37); P < 0.0001, and -0.54 (-0.93, -0.14); P = 0.0079, respectively. Adverse events were consistent with current product labeling, with dizziness the most commonly reported adverse event (24.3% of pregabalin-treated patients). ConclusionPregabalin improved measures of pain and sleep, and is well tolerated in Chinese patients with PHN. These results may inform physicians treating patients with PHN in China.
Background: Necrotizing enterocolitis (NEC) is the leading cause of neonatal gastrointestinal mortality; effective interventions are lacking with limited understanding of the pathogenesis of NEC. The importance of Toll-like receptor 4 (TLR4) signaling in NEC is well documented; however, the potential mechanisms that regulate enterocyte inflammation and apoptosis remain unclear. The aim of this study was to characterize the role of TLR4-mediated inflammation and apoptosis in the development of NEC and to determine the major apoptotic pathways and regulators in the process. Methods: TLR4-deficient C57BL/10ScNJ mice and lentivirus-mediated stable TLR4-silent cell line (IEC-6) were used. NEC was induced by formula gavage, cold, hypoxia, combined with lipopolysaccharide in vivo or lipopolysaccharide stimulation in vitro. Enterocyte apoptosis was evaluated by TUNEL or Annexin analysis. The expression of TLR4, caspase3, caspase8, caspase9, Bip, Bax, Bcl-2, and RIP was detected by Western blot and immunofluorescence. Inflammatory factors such as tumor necrosis factor-a and interleukin-2 were examined by Luminex. Results: Defect of TLR4 led to suppressed enterocytes apoptosis both in vitro and in vivo; the expression of caspase3, caspase8, Bip, and Bax was decreased; and caspase9 and Bcl-2 were increased. NEC severity was attenuated in TLR4-deficient mice compared with wild-type counterparts, and enterocytes apoptosis was correlated with NEC severity. RIP and cytokine level of tumor necrosis factor-a and interleukin-2 were also decreased. Conclusions: TLR4-induced inflammation and apoptosis play a critical role in the pathogenesis of NEC. TLR4 inhibition, combined with extrinsic (caspase8) and/or endoplasmic reticulum stress (Bip) apoptosis signaling blockade could serve as a potential effective treating strategy for NEC.
Rationale: Ebstein anomaly is a common congenital heart disease that may induce severe tricuspid regurgitation and dilation of the "atrialized" portion of the right ventricle. Patients who undergo surgery to correct Ebstein anomaly are at high risk of postoperative right-sided heart failure, yet little is known about what pre-, peri-, or postoperative procedures may help reduce this risk. Patient concerns: Here, we describe 3 cases of adults with Ebstein anomaly who underwent corrective surgery and in whom right-sided heart failure occurred with severe tricuspid regurgitation detected by transesophageal echocardiography. Diagnoses: Ebstein anomaly. Intervention: Various approaches were applied to prevent right heart failure: perioperative control of atrial and ventricle arrhythmia, protection of myocardium, reduction of right-side cardiac workload after cardiopulmonary bypass, and mechanical support for right heart. Outcomes: One of the 3 patients died, another experienced kidney failure despite postoperative support on extracorporeal membrane oxygenation, and the third patient survived without complications. Lessons: Our case series suggests that surgical prognosis can be improved through aggressive preoperative treatment, vasoactive and anti-arrhythmia medications, and comprehensive measures designed to reduce myocardial injury, prevent myocardial edema, and reduce pre- and afterload on the right ventricle.
目的：了解大鼠脑出血后血肿周围组织细胞凋亡与神经元特异性烯醇化酶（NSE）的表达及大鼠神经功能缺损程度的关系。方法：用胶原酶注入到大鼠尾状核的方法制作脑出血模型。将大鼠分为脑出血、假手术组、正常组3组。采用苏木素伊红(HE) 染色、NSE免疫组织化学染色及TUNEL分别观察各组在脑出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d时血肿周围NSE及TUNEL的表达。用Longa评分法评价大鼠神经功能缺损程度。结果：大鼠在胶原酶注入6 h后形成稳定的血肿，在造模24～48 h神经功能缺损程度最重；6 h即见到TUNEL阳性细胞的表达，在48 h最明显；NSE从神经元中漏出弥散到细胞间隙也在48 h达高峰。结论：脑出血血肿周围凋亡与神经功能缺损及NSE的变化有关，凋亡可能在脑出血的神经损伤中起重要的作用。
Interleukin-10 (IL-10)-producing B cells (B10 cells) play a critical role in the immune system balance by negatively regulating inflammatory responses. This study was conducted to determine the effect of local B10 cell induction on periodontal inflammation and bone loss in ligature-induced experimental periodontitis in vivo. Purified spleen B cells from C57BL/6J mice (8 to 10 weeks old) were cultured with CD40 ligand (CD40L) and the Toll-like receptor 9 (TLR9) agonist cytidinephosphate- guanosine oligodeoxynucleotide (CpG) to determine effective IL-10 induction in vitro. Silk ligatures (size 7-0) were tied around the mouse maxillary second molars on day 0, followed by the injection of CD40L and CpG into the palatal gingiva on days 3, 6, and 9. All the mice were sacrificed, and samples were collected on day 14. CD40L and CpG significantly increased the level of IL-10 production by B cells in vitro, although the frequencies of CD1(dhi) CD5 (+) and IL-10-producing (IL-10 (+)) CD45 (+) cells were decreased. IL-10 was predominantly produced by the CD1(dhi) CD5 (+) subpopulation of B cells. In vivo, both IL-10 mRNA expression and the number of IL-10 (+) CD45 (+) cells were significantly increased after gingival injection of CD40L and CpG. Periodontal bone loss was significantly decreased and the gingival expression of IL-1 (+), tumor necrosis factor alpha, and RANKL was significantly reduced. The number of multinucleated tartrate-resistant acid phosphatase-positive cells along the alveolar bone surface was significantly decreased after gingival injection of CD40L and CpG. This study indicates for the first time that the local induction of B10 cell activity could inhibit periodontal inflammation and bone loss.
Aim: Inflammation plays a role in secondary brain injury after intracerebral hemorrhage (ICH). We aimed to determine the prognostic significance of admission white blood cell (AWC), neutrophil count (ANC), lymphocyte count, monocyte count and neutrophil to lymphocyte ratio (NLR) for 90-day outcome after ICH. Patients & methods: A total of 336 patients with spontaneous ICH were retrospectively investigated. Clinical outcome was assessed by modified Rankin Scale at 90 days. Results: Multivariate analysis showed that higher AWC, ANC, NLR were independently associated with mortality and worse outcome. Moreover, NLR showed a higher predictive ability in mortality than in poor outcome in receiver operating characteristic analysis. Linear regression analyses revealed admission Glasgow Coma Scale score and ICH volume were mostly correlated with these indices. Conclusion: Elevated levels of AWC, ANC and NLR were independently related to poor 90-day outcome after ICH. NLR may be a novel inflammatory biomarker following ICH.
The plasminogen activator inhibitor-1 (PAI-1) is a candidate gene for stroke based on PAI-1's crucial role in fibrinolytic system. However, association studies have yielded conflicting results regarding the association between PAI-1 polymorphisms and stroke susceptibility. To further elucidate the putative association, we performed a systematic review and meta-analysis to provide a complete picture of the loci investigated for association of PAI-1 polymorphism with stroke risk and to derive a precise estimation. PubMed, Embase, and China National Knowledge Infrastructure (CNKI) databases were searched until June 2015 to identify eligible studies. Forty data sets from 39 studies with a total of 8336 cases and 14,403 controls were included. The most commonly investigated polymorphism was -675 4G/5G, followed by -844 G/A, 11053 T > G, HindIII C/G, and (CA)n. Overall, our meta-analysis provided evidence for the significant association of PAI-1 4G/5G polymorphism with an increased risk of adult but not pediatric ischemic stroke (adult: 4G/4G vs. 4G/5G + 5G/5G, OR = 1.21, 95 % CI = 1.04-1.42). In the subgroup analysis, significant association was detected in Asians (4G/4G vs. 4G/5G + 5G/5G, OR = 1.45, 95 % CI = 1.14-1.85) but not Caucasians. Moreover, we found that -844 G/A but not 11053 T > G polymorphism was associated with an increased risk of ischemic stroke (-844G/A: A/A vs. G/G: OR = 1.32, 95 % CI = 1.01-1.73). A tendency of PAI-1 4G/5G polymorphism towards a decreased risk of hemorrhagic stroke was observed (4G/4G + 4G/5G vs. 5G/5G, OR = 0.77, 95 % CI = 0.59-1.02, P = 0.066). Future well-designed studies in large well-characterized sample size and presenting results stratified by gender, age, and stroke subtype are warranted.
摘要:目的:分析高胆红素血症新生儿血清神经元特异性烯醇化酶(NSE)含量和新生儿行为神经能力测评(Neonatal Behavioral Neurological Assessment,NBNA)的变化,探讨高胆红素血症新生儿血清NSE含量变化的临床意义。方法:应用放射免疫分析法分别测定60例高胆红素血症新生儿和20例对照组新生儿血清NSE含量,同步测定血清总胆红素(TSB),进行NBNA评分;高胆红素血症组早期干预后再次测定血清NSE含量。结果: 与对照组比较,高胆红素血症新生儿血清TSB、NSE含量显著升高,而NBNA评分明显降低,差异有显著性意义(Plt;0.01);对照组与高胆红素血症新生儿轻度增高、中度增高、重度增高四组两两比较(均Plt;0.05),存在显著性差异;血清NSE含量与NBNA评分呈明显负相关(r=-0628,Plt;0.01);高胆红素血症新生儿经早期干预治疗后,血清NSE含量均下降(Plt;0.05),差异有显著性。结论: 高胆红素血症可导致新生儿脑损伤,血清NSE含量可以作为脑损伤的监测指标。Abstract: Objective: To analyze levels of neuronspecific enolase(NSE)in serum and neonatal behavioral neurological assessment (NBNA), to study whether NSE in serum can be used as a tool for the early identification of brain damage in neonatal hyperbilirubinemia. Methods: Serum NSE level of 60 full term infants with hyperbilirubinemia and 20 cases as to control group were measured by radioimmunoassay; Also total serum bilirubin (TSB) and NBNA were detected. In the hyperbilirubinemia group,serum NSE level were measured second when TSB were less than 855 μmol/L(5 mg/dL). Results: Compared with control group,the levels of serum TSB、NSE of the hyperbilirubinemia group were significantly higher, but NBNA score was significantly lower. The levels of serum NSE was significantly negative related to NBNA score. In the hyperbilirubinemia group, serum NSE level were significantly lower after treatment. Conclusion: Hyperbilirubinemia in neonates can cause brain damage. Serum NSE level could work as monitoring indexes of this damage.