清解防感颗粒体内外抑菌作用实验研究

目的 :研究清解防感颗粒的体内外抑菌作用。方法 :采用标准试管二倍稀释法测定清解防感颗粒的最低抑菌浓度(MIC)和平板转种法测定最低杀菌浓度(MBC),并观察其在小鼠体内外对金黄色葡萄球菌、表皮葡萄球菌和乙型溶血性链球菌的抑菌作用。结果 :体外抑菌试验显示:清解防感颗粒对上呼吸道常见细菌有不同程度的抑菌作用,其中对金黄色葡萄球菌、表皮葡萄球菌和乙型溶血性链球菌的作用较强,对铜绿假单孢菌和白色念珠菌也有一定的抑制作用。体内抑菌作用显示:清解防感颗粒8、4 g/kg剂量组有抑菌作用,可明显降低金黄色葡萄球菌感染小鼠的死亡率并延长生存期。结论 :清解防感颗粒具有一定的体内外抑菌作用。     

IMB模型的理论框架与运用

<正>IMB模型(信息-动机-行为技巧模型,Information-Motivation-Behavioral Skills Model,简称IMB)由JD费舍尔和WA费舍尔在1992年最早提出,与以往的健康相关行为改变模式相比,将影响行为改变的各种因素分为信息、动机、行为技巧3个组分,并验证自身具有的预防疾病的信息和参与预防的动机可通过行为技巧引发预防行为的启动和维     

丹参多酚酸与三七总皂苷合用保护OGD/R损伤胶质细胞线粒体、促进神经因子表达作用研究*

目的 探讨丹参多酚酸（Salvianolate Acid，SAL）和三七总皂苷（Panax Notoginseng Saponins，PNS）合用保护氧糖剥夺/复氧复糖（Oxygen-Glucose Deprivation/Reoxygenation，OGD/R）损伤后星形胶质细胞线粒体、促进神经营养因子表达的作用及机制。方法 大鼠星形胶质细胞原代培养，建立OGD/R损伤模型。Cell counting kit-8（CCK-8）法检测星形胶质细胞活力，流式细胞仪检测活性氧（reactive oxygen species，ROS）释放量、胞内Ca2+ 浓度、线粒体膜电位（mitochondrial membrane potential，MMP）变化，实时定量PCR（real time polymerase chain reaction，RT-PCR）和蛋白质免疫印迹（western blot，WB）法检测HIF-1α、PI3K/Akt/mTOR信号通路蛋白及脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）、神经生长因子（nerve growth factor，NGF）、胰岛素样生长因子（insulin-like growth factors，IGF-1α）的表达情况。结果 胶质细胞OGD/R条件确定为OGD4h/R24h，给药浓度确定为SAL 25 μg·mL-1、PNS 6.25 μg·mL-1。与OGD/R组相比，SAL 25 μg·mL-1、PNS 6.25 μg·mL-1、SAL 25 μg·mL-1与PNS 6.25 μg·mL-1合用均能增强损伤后星形胶质细胞活力（P＜0.05），升高MMP，降低ROS释放量（P＜0.05），降低损伤后星形胶质细胞HIF-1α蛋白和mRNA的表达，升高磷脂酰肌醇激酶（PI3K）磷酸化水平（P＜0.05）；SAL还可升高mTOR蛋白磷酸化水平（P＜0.05），PNS可提高Akt磷酸化水平（P＜0.05），增加损伤星形胶质细胞BDNF mRNA 表达（P＜0.05）；SAL、SAL与PNS合用能增加损伤后星形胶质细胞 BDNF、NGF、IGF-1α mRNA 表达（P＜0.05）。结论 丹参多酚酸和三七总皂苷组分合用可保护OGD/R损伤后星形胶质细胞线粒体、促进胶质细胞表达神经营养因子BDNF、NGF、IGF-1α。     

巨细胞病毒pp65抗原检测的新方法

目的传统的检测CMVpp65抗原的方法是间接免疫荧光,操作麻烦,步骤较多,整个实验需要6个多小时,而且只能定性。需要寻找一种简便的可以定量的方法来检测CMVpp65抗原阳性细胞。因此我们设计了流式的方法来检测CMVpp65抗原阳性细胞。方法我们用间接免疫荧光试剂盒中的一抗和购自invitrogen公司的二抗进行了流式标记移植病人外周血中的WBC,操作步骤明显简便了,只需不到2 h就可完成检测,而且可以定量检测CMVpp65抗原阳性细胞。同时我们用荧光定量PCR检测病人尿中的CMV基因的拷贝数。结果流式检测阳性率高的标本,用间接免疫荧光方法检测也为阳性,同时流式检测强阳性的病人尿用荧光定量PCR检测到CMV基因,表明病人处于CMV病毒血症。结论我们用间接免疫荧光试剂盒中的一抗和购自invitrogen公司的二抗进行流式标记,成功检测了移植病人的CMVpp65抗原阳性细胞的百分比,新方法可以定量,与荧光定量PCR方法检测病人尿中的CMV基因的结果符合率较高,可以推广应用。     

Revisiting colorectal cancer animal model – An improved metastatic model for distal rectosigmoid colon carcinoma

Colorectal cancer (CRC) is the second most frequent and fatal cancer in Western countries. Understanding its biology with different incidence along the colon and rectum, genetic profile and how these factors contribute to local/distant progression, has been hampered by the lack of a suitable CRC model.We report a reproducible model, using human CRC cell lines (CL) (WiDr, LS1034, C2BBe1) injected (1?×?10⁷ cells/animal) in RNU rats (n?=?55) which underwent cecostomy and descending colostomy with mucosal-cutaneous fistula of the sigmoid colon. CL were characterized by immunohistochemistry: CK20, CDX2, P53, vimentin, Ki67, CD44, CD133, E-cadherin, β-catenin and CEA; cancer stem cells-immune system interaction was studied and tumor progression was assessed with nuclear medicine imaging (^99mTc-MIBI).Animals developed locally invasive tumors and with WiDr neural invasion was registered. Cancer stem cells were detected in WiDr (CD44 positive). All the cell lines stimulated the immune system, being WiDr the most aggressive. Imaging studies demonstrated tumor uptake.With this CRC model we can study the microenvironment role and tumor-stroma interactions. All CL developed primary disease, but only the WiDR established neural invasion which may represent a metastatic pathway. This model can help unveiling the underlying metastatic mechanisms, and ultimately test better therapeutic approaches for CRC.     

老年慢性阻塞性肺疾病急性加重期患者IL-8、IL-6、TNF-α水平变化与肺功能的相关性研究

目的探讨老年慢性阻塞性肺疾病急性加重期(AECOPD)患者IL-8、IL-6、TNF-α水平变化和肺功能的相关性。方法选取我院收治的AECOPD患者78例。分别于患者急性加重期和稳定期检测其血清IL-8、IL-6、TNF-α水平以及肺功能。对比不同肺功能综合评估分级的患者IL-8、IL-6、TNF-α的水平,同时对比急性加重期和稳定期患者IL-8、IL-6、TNF-α和FEV1%预计值的检测结果,并分析IL-8、IL-6、TNF-α与FEV1%预计值的相关性。结果肺功能综合评估分级级别越高,其IL-8、IL-6和TNF-α的水平显著增高(p<0.05);COPD急性加重期及稳定期患者的IL-8、IL-6、TNF-α水平和FEV1%预计值水平均呈负相关(p<0.05)。结论老年AECOPD患者IL-8、IL-6和TNF-α水平和FEV1%预计值呈负相关,检测IL-8、IL-6和TNF-α的水平可反应出患者病情的严重程度,有助于临床评估患者的病情,指导临床治疗。     

Expression and function of Erbin protein in renal ischemia-reperfusion injury

Objective To investigate the expression of ErbB2 interacting protein (Erbin) in renal ischemia-reperfusion injury (IRI) in vivo and in vivo, and the effect of Erbin over-expression on IRI. Methods (1) In vivo, the model of renal IRI was constructed in mice, and set up sham group and reperfusion 3, 6, 12, 24 and 48 h IRI group. BUN and Scr were detected and PAS staining was used to observe the pathology change of renal tissues. Cell apoptosis was detected by TUNEL staining. Erbin and NF-κB expression in renal tissue was detected by Western blotting, and immunohistochemistry was used to detect the distribution of Erbin. (2) In vivo, IRI model in HK2 cells was constructed and cells were harvested after culturing in normal medium for 3, 6, 12 and 24 h. Erbin expression was detected by Western blotting. Flow cytometry and ELISA were used to evaluate the level of cell apoptosis and inflammatory cytokine secretion respectively. HK2 cells were transiently transfected with Prk5-myc-Erbin plasmid via lipofectamine 2000, and were divided into control group, IRI group, Erbin group and Erbin+IRI group. The protein expression of Erbin and NF-κB, cell apoptosis and inflammatory cytokine secretion was detected. Results (1) Compared with sham group, serum BUN and Scr were dramatically increased in IRI model, especially in 24 h after reperfusion (P＜0.05). Moreover, PAS staining showed that a lot of renal tubular epithelial cells were necrosis and fell off, and many protein cast were formed, renal injury score and apoptotic index were higher in 6 h, 12 h, 24 h, 48 h IRI model than those in sham group (all P＜0.05). The expression of Erbin, which was expressed in renal tubules, and nuclear NF-κB in 24 h IRI model were significantly increased, as compared with sham group (all P＜0.05). (2) Compared to those in control group, nuclear NF-κB expression, apoptosis and inflammatory cytokine secretion were significantly increased in IRI group. Meanwhile, Erbin expression was also induced and peaked at 24 h (P＜0.05). Compared to those in IRI group, cell apoptosis, the expression of nuclear NF-κB, inflammatory cytokine IL-6 and TNF-α were decreased in Erbin+IRI group (all P＜0.05). Conclusions Erbin expression is up-regulated in renal IRI, and over-expression of Erbin can partly inhibit NF-κB activation, cell apoptosis and inflammatory cytokine secretion in IRI group, which indicates Erbin may playing a protective role in renal IRI.     

中老年急性肺血栓栓塞症的诊治分析

目的探讨1996～2010年江苏省人民医院确诊为急性肺血栓栓塞症（APTE）中老年病例的危险因素、易患因素以及诊断和治疗方法的变迁。方法对我院1996～2010年间280例确诊为APTE的中老年病例的发病特点、诊治及其转归进行临床回顾性评估分析。结果 APTE好发年龄为50～80岁,占发病的77.5%,以冬春季节为多。APTE临床表现仍以胸闷气急、呼吸困难为主要临床表现,占60.7%。D二聚体〉0.5μg/L占88.5%,仍是高敏感性检查指标。结论 APTE发病与易发因素有关,对其认识加深,重视高敏感性检查指标的筛选,结合快速有效的影像学检查,可提高APTE的诊断率。