Transduction of avian cells with recombinant baculovirus

Transduction of chicken and duck cells was examined by Ac-CMV-eGFP, a recombinant baculovirus capable of expressing an eGFP reporter gene under the control of the CMV promoter. The results showed that chicken and duck cells were transducible, as demonstrated by a flow cytometry assay. The transduction efficiency of duck cells was higher than that of chicken cells. The addition of histone deacetylase inhibitor sodium butyrate enhanced the expression levels of eGFP both in chicken and duck cells. Ac-CMV-eGFP is capable of transducing genes into a variety of chicken cells in organs such as liver, lung and kidney. Of three cells from different organs tested, the highest transduction was observed in lung cells (49.8%), followed by kidney cells (44%) and liver cells (43%). Only in chicken liver cells AcMNPV transduction was in a dose-dependent manner. It also showed that baculovirus enters the avian cells by endocytosis and is released into the cytoplasm by acid-induced fusion.     

Location and phylogenetic analysis of the region immediately upstream of the granulin gene of the Clostera anachoreta granulovirus

The region immediately upstream of the granulin gene from Clostera anachoreta granulovirus (ClanGV) was identified from hybridization experiments and sequenced. The sequence of 5122nt EcoRI restriction fragment was presented and compared with the equivalent area in other GVs. Database searches showed that this region contained three open reading frames (ORFs) similar to the baculovirus genes (egt, fgf and me53, respectively) and four ORFs unique to ClanGV genome. Phylogenetic trees of the baculovirus genes egt and me53 were constructed. These analyses indicated that ClanGV genes may be more closely related to CfGV, CpGV, ClGV, PoGV and AoGV than to PxGV and XcGV.     

Serum CYFRA 21-1 level reflects hepatocellular carcinoma metastasis: study in nude mice model and clinical patients

Our previous proteomics study on human hepatocellular carcinoma (HCC) cell strains revealed that cytokeratin 19 (CK19) was expressed in cells with high metastasis potential; we further studied serum CK19 fragment CYFRA 21-1 level in HCC patients and nude mice model of HCC metastasis. HCC cell line HCCLM3 was injected subcutaneously into 30 nude mice which were then randomized into 6 groups of 5 mice each. The murine serum CYFRA 21-1 and pulmonary metastases were determined 2, 3, 4, 5, 6, and 7 weeks after injection. Serum CYFRA 21-1 levels of 101 normal controls and 108 HCC patients were also determined. In nude mice model, CYFRA 21-1 level increased significantly when pulmonary metastases occurred. Among 108 HCC patients, 24 (22.2%) had increased serum CYFRA 21-1 level. The presence of portal vein tumor emboli was significantly higher in CYFRA 21-1 increased cases (33.3%, 6/24) than in CYFRA 21-1 normal cases (6.0%, 5/84) (x ² = 7.403, P < 0.01). In addition, the percentage of TNM stage III/IV tumor was significantly higher in CYFRA 21-1 increased patients (54.2%, 13/24) than in CYFRA 21-1 normal cases (21.4%, 18/84) (x ² = 9.776, P < 0.005). These results suggest that CK19 may play an important role in HCC metastasis.     

醛固酮对体外新生大鼠心肌成纤维细胞增殖及胶原代谢的影响

目的:探讨醛固酮对体外新生大鼠心肌成纤维细胞(CFs)增殖及胶原代谢的影响并探讨其机制。方法:体外差速贴壁法分离纯化培养CFs,试验分为4组:对照组、醛固酮组、醛固酮+螺内酯组、醛固酮+洛沙坦组。各组作用24h后,MTT法检测CFs增殖;流式细胞分析仪分析细胞周期;分光光度法检测细胞上清中羟脯氨酸含量;免疫细胞化学荧光染色法检测纤维连接蛋白(FN)及增殖细胞核抗原(PCNA)的蛋白表达。结果:与对照组比较,醛固酮组可显著增强细胞活性,使S期细胞百分率增加,G0/G1期细胞百分率下降,羟脯氨酸含量增加,PCNA、FN的蛋白表达增强(P<0.01)。与醛固酮组比较,醛固酮+螺内酯组的CFs增殖及胶原合成被抑制,PCNA、FN的蛋白表达下降(P<0.05),醛固酮+洛沙坦组抑制CFs的增殖及胶原合成,FN的蛋白表达降低(P<0.01),而PCNA的蛋白表达无明显改变(P>0.05)。结论:醛固酮通过其受体及AT1受体途径促进CFs增殖及胶原合成,且醛固酮受体通路占主导地位;其作用机制可能与促进PCNA、FN的蛋白表达有关。     

Anaplastic myeloma and T-cell lymphoma in the same patient

To the Editor:Anaplastic myeloma is a rare variety of multiple myeloma and the coexistence of anaplastic myeloma and peripheral T cell lymphoma has never been reported. A 48-year-old woman admitted to the hospital with no obvious cause of fever (FUO) for two weeks.The body temperature rose in the afternoon and at night,with peak temperature over 40℃.Moreover,the pre-hospital anti-inflammatory therapy was ineffective.Physical examination revealed no apparent abnormality.The leukocyte count was 86× 109/L with 60.6％ neutrophils and 30％ lymphocytes which did not support the existence of bacterium or virus infection.The tests for parasite,mycoplasma,chlamydia and tuberculosis were all negative.Furthermore,ultrasonography of the abdomen and the test of distinctive tumor markers showed no abnormalities.However,the bone marrow cytomorphological examination showed that the plasma cells were up to 14％ and the immature plasma cells were about 9.5％.     

Current role of surgery for the treatment of fulminant Clostridium difficile colitis

Objective This review discusses the current status and progress in studies on fulminant Clostridium difficile colitis (FCDC),including the definition,risk factor,diagnostic role of CT,surgical treatmen...     

谁之正义:中国公立医院改革研究管见

围绕中国公立医院改革而展开的学术研究活动经历了三个阶段:1997～2005年是酝酿和前期试验期;2006～2009年为"探索等待期";2010至今为研究高峰期。学术研究活动对改革实践的促进作用体现在思想启蒙、宣传教育、交流信息、开阔思路等方面,取得的成绩有目共睹。其不足之处在于,研究者对改革模式的设计和思考漠视患方利益,对新格局下的卫生规划与布局缺乏前瞻性和必要的人文意识,研究对国家相关卫生政策亦步亦趋,缺乏足够的理论勇气和创新动力。     

Assessment effect of central venous pressure in fluid resuscitation in the patients with shock: a multi-center retrospective research

Background Central venous pressure (CVP) and intrathoracic blood volume index (ITBVI) were used to assess the fluid status.It has previously been shown that CVP is not as accurate as ITBVI for all the shock patients.We therefore hypothesized that the change of CVP has the ability to predict fluid responsiveness in some clinical cases of shock.Methods From September 1st 2009 to September 1st 2011,sixty-three patients with shock from different Intensive Care Unit (ICU) were collected into this retrospective study.All the patients received fluid challenge strategy (infusing 300 ml hydroxyethyl starch in 20 minutes),were monitored with CVP and pulse-indicated continuous cardiac output (PICCO).The correlation between changes in cardiac index (△CI),CVP (△CVP) and ITBVI (△ITBVI) were analyzed.Fluid responsiveness was defined as an increase in CI≥10％.Receiver operating characteristic (ROC) curves were generated for △CVP and △ITBVI.Results For all the patients,there was no correlation between △CI and △CVP (P=0.073),but in the subgroup analysis,the correlation between △CI and △CVP was significant in those younger than 60 years old (P=-0.018) and those with hypovolemic shock (P=0.001).The difference of areas under the ROC curves of △CVP and △ITBVI were not statistically significant in the group younger than 60 years old or hypovolemic shock group (P ＞0.05,respectively).However,no similar results can be found in the group older than 60 years old and the other two shock type groups from ROC curves of △CVP and △ITBVI.Conclusions △CVP is not suitable for evaluating the volume status of the shock patients with fluid resuscitation regardless of their condition.However,in some ways,△CVP have the ability to predict fluid responsiveness in the younger shock patients or in the hypovolemic shock patients.     

Statins protect human endothelial cells from TNF-induced inflammation via ERK5 activation

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) exert pleiotropic effects on the cardiovascular system, in part through a decrease in reactive oxygen species (ROS) formation and reduction of vascular inflammation. To elucidate the molecular mechanisms involved in these effects, we investigated the effect of statins on TNF-α-induced ROS production, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression in human aortic endothelial cells (HAECs). Exposure of HAECs to TNF-α caused production of ROS via Rac-1 membrane translocation and activation. The Rac-1 activation and ROS liberation mediated TNF-stimulated NF-κB activation and the subsequent VCAM-1 and ICAM-1 expression. Extracellular-signal-regulated kinase 5 (ERK5) plays a central role in inhibiting endothelial inflammation. Immune complex kinase assay of protein extracts from HAECs treated with atorvastatin revealed increased ERK5 activity in a time- and dose-dependent manner. In addition, pretreatment with atorvastatin inhibited TNF-α-induced ROS production and VCAM-1 and ICAM-1 expression. Chemical or genetic inhibition of ERK5 ablated the statins inhibition of Rac-1 activation, ROS formation, NF-κB, VCAM-1 and ICAM-1 expression induced by TNF-α. Taken together, statins, via ERK5 activation, suppress TNF-stimulated Rac-1 activation, ROS generation, NF-κB activation and VCAM-1 and ICAM-1 expression in human ECs, which provides a novel explanation for the pleiotropic effects of statins that benefit the cardiovascular system.     

Comprehensive Study of the Interaction Between a Potential Antiprion Cationic Porphyrin and Human Prion Protein at Different pH by Using Multiple Spectroscopic Methods

Cationic porphyrins are potential antiprion drugs; however, the action mechanisms remain poorly understood. Herein, the interaction between a cationic porphyrin and recombinant human prion protein (rPrP^C) was comprehensively studied by using surface plasmon resonance (SPR), fluorescence, resonance light scattering (RLS), and circular dichroism (CD) spectroscopy. The experimental results showed that the interaction between the cationic porphyrin and rPrP^C was pH dependent. The equilibrium association constants obtained from SPR spectroscopy were 4.12 × 10³ M^? 1 at pH 4.0, 1.74 × 10⁵ M^? 1 at pH 6.0, and 5.98 × 10⁵ M^? 1 at pH 7.0. The binding constants at 298 K obtained from the fluorescence quenching method were 7.286 × 10⁴ M^? 1 at pH 4.0 and 1.45 7 × 10⁵ M^? 1 at pH 6.0. The thermodynamic parameters such as enthalpy change, entropy change, and free energy change were calculated, and the results indicated hydrogen bonds and van der Waals interactions played a major role in the binding reaction. The RLS experiment was performed to study the influence of porphyrin on the rPrP^C aggregation at different pH values. The CD experiments were conducted to investigate the effects of porphyrin on the secondary structure and thermal stability of rPrP^C. Finally, the comparison of SPR measurement and fluorescence quenching measurement was discussed.