大剂量HBsAg疫苗对HBV转基因小鼠细胞免疫调节的研究

目的　研究大剂量HBsAg疫苗对HBV转基因小鼠 (Tg鼠 )细胞免疫应答的影响。方法　Tg鼠 4 8只 ,每只注射 6 μg(相当于人产生抗体剂量的 180倍 )的HBsAg疫苗后 ,用流式细胞术、氚 胸腺嘧啶核苷 (3 H TdR)掺入法和ELISA法 ,分别检测其树突状细胞(DC)、T淋巴细胞增殖及其诱生细胞因子IL 2、IFN γ的水平。结果　大剂量HBsAg疫苗组DC表面共刺激分子CD80、CD86和I Ek的阳性百分率 (% )均高于对照组 (P <0 0 5 ) ,特异T淋巴细胞增殖能力 8个月的cpm值 (10 0 77 2± 85 74 0 )显著高于 1周的 (5 32 9 1± 30 86 0 ) (P <0 0 5 ) ;其诱生细胞因子的水平 8个月时IL 2 (46 2 4± 12 2 5pg/ml)、IFN γ(976 1± 5 4 4 1pg/ml)显著高于 1周时IL 2 (15 6 1±78 7)、IFN γ(5 8 3± 4 9 5 ) (P <0 0 1)。结论　大剂量HBsAg疫苗能够增强Tg鼠细胞免疫应答能力 ,恢复对乙型肝炎抗原的应答。     

干扰素-γ对动脉损伤后平滑肌细胞增殖和迁移的作用

目的　探讨干扰素 γ(IFN γ)对动脉损伤后平滑肌细胞 (SMC)增殖和迁移的作用及其作用机制。方法　建立兔髂动脉球囊损伤动物模型 ,培养损伤后动脉SMC ,实验分四组 :对照组、转化生长因子 β1(TGF β1)组、IFN γ组、TGF β1+IFN γ组。每组细胞加培养液或加入 10ng/mlTGF β1或 /和5 0 0U/mlIFN γ刺激 72h ,细胞计数和四甲基偶氮唑盐 (MTT)法测定各组SMC的增殖 ,同时测定SMC迁移。明胶酶谱分析SMC中MMP 2的活力。结果　与对照组比较 [细胞数 (2 .875± 0 .32 3)× 10 5个 /ml,迁移距离 (2 79.9± 8.12 9) μm],72h后TGF β1组细胞数 (4 .188± 0 .2 39)× 10 5个 /ml,(P <0 .0 1)和迁移距离 (36 5 .8± 9.6 86 ) μm ,(P <0 .0 1)增加 ,MTTA值检测细胞增殖抑制率为 - 19.4 % ;IFN γ组细胞数 (1.938± 0 .2 4 9× 10 5个 /ml,P <0 .0 1)和迁移距离 (2 34.4± 9.72 2 μm ,P <0 .0 1)减少 ,细胞增殖抑制率为 15 .8% ;TGF β1+IFN γ组细胞数 (3.12 5± 0 .2 5 4× 10 5个 /ml,P <0 .0 1)和迁移距离(32 3.1± 8.4 81) μm ,(P <0 .0 1)低于TGF β1组 ,细胞增殖抑制率为 - 9.1%。明胶酶谱分析显示各组细胞均可检测到基质金属蛋白酶 2 (matrixmetalloproteinase 2 ,MMP 2 6 6× 10 3 u) ,TGF β1组还可检     

CIK细胞的表型分析及生物学活性研究

目的　体外诱导培养细胞因子诱导的杀伤细胞 (CIK) ,并研究其表型及生物学活性。方法　从外周血分离淋巴细胞 ,经过IFN γ、CD3McAb及IL 2诱导并培养 ,获得大量的CIK细胞。FACS测定CIK表面CD3CD5 6、CD3、CD5 4、HLA DR、CD11a、CD2 8、CD86、CD80等CD分子表达情况及其百分率 ,3 H TdR检测其增殖能力 ,MTT法检测CIK对胃癌MGC 80 3细胞、肝癌Bel 74 0 2细胞的杀伤活性。结果　CIK细胞高度表达CD3、CD5 4、CD11a,中度表达CD3CD5 6、HLA DR、CD2 8CD5 4、CD2 8,不表达CD86、CD80 ,且具有较强的增殖能力及非MHC限制性杀瘤活性。结论　IFN γ、CD3McAb及IL 2三种细胞因子体外诱导单个核细胞可获得具有高增殖活性及较强杀瘤活性的CIK细胞     

强直性脊柱炎患者外周血单个核细胞干扰素诱导蛋白10及Th1/Th2型细胞因子的表达

目的　探讨干扰素诱导蛋白 10 (IP 10 )在强直性脊柱炎 (AS)发病中的作用。方法　收集肝素抗凝的AS患者的外周全血 ,梯度离心得到外周血单个核细胞(PBMC) ,应用半定量RT PCR法测定IP 10、IFN γ和IL 4mRNA表达水平。结果　AS患者PBMC中IP 10、IFN γ和IL 4mRNA表达水平均较健康对照组高(P <0 0 1) ,同时IFN γ/IL 4值也较对照组高 (P <0 0 5 )。IFN γ和IL 4mRNA表达水平与C 反应蛋白呈负相关 ,IP 10mRNA表达水平与IFN γ、IL 4mRNA和IFN γ/IL 4值均呈正相关。结论　AS患者PBMC表达的细胞因子以Th1型为主 ,IP 10表达升高 ,IP 10可能通过促进Th1型细胞向炎症部位的归巢来参与AS的发病     

哮喘患儿T细胞亚群功能状态的研究

为观察哮喘患儿T细胞亚群功能状态 ,以探讨其发病机制 ,分别采用酶联免疫吸附试验 (ELISA)、流式细胞术 (FCM)检测哮喘患儿外周血单个核细胞 (PBMC)培养上清液中IL 4、IFN γ及PBMC细胞膜上CD30的表达。结果显示 ,发作组PBMC培养上清液中IFN γ为 338 98±90 4 9ng/L ,IL 4为 37 75± 1 1 94ng/L ,IgE为 2 82 39± 83 33U/L ,与对照组比较差异有显著性意义 (P均 <0 0 1 ) ;经治疗后IFN γ为 4 0 8 6 9±93 5 8ng/L ,IL 4为 33 35±1 1 99ng/L ,IgE为 1 37 2 8± 89 0 9U/L ,与对照组比较差异有非常显著性意义 (P均 <0 0 1 ) ;IL 4与IgE呈正相关关系(r=0 5 6 1 ,P <0 0 5 ) ;哮喘患儿PBMCCD30表达增强。提示哮喘患儿存在T细胞亚群功能紊乱 ,Th1 /Th2的失衡在哮喘发病机制中起着很重要的作用     

严重急性呼吸综合征患者细胞因子和外周血淋巴细胞的动态变化

目的　观察严重急性呼吸综合征 (SARS)患者外周血淋巴细胞绝对细胞数和细胞因子在发病过程中的动态变化 ,了解SARS患者机体的免疫状态。方法　用酶联免疫吸附法 (ELISA)动态检测 2 1例SARS患者发病过程中血浆细胞因子IFN α、TNF α、IFN γ、IL 12和IL 10的水平。结果　分别有 6 6 7%的患者外周血淋巴细胞绝对细胞数、38 1%的患者血浆IFN α水平和 6 1 9%的患者血浆TNF α水平在发病中期达到高峰。部分患者Th1细胞因子IL 12和IFN γ有所升高 ,而所有患者整个检测时间内Th2细胞因子IL 10的产生未受到影响。 85 7%的患者发病后血浆IFN α的首次检测值 (16 1 0 9± 132 6 9pg/ml)显著低于正常值 (86 7 18± 30 6 5 0 pg/ml,P <0 0 5 )。结论　SARS病毒感染后机体的免疫反应在病程中期可能最强 ,且可能以Th1反应为主。血浆IFN α首次检测值显著低于正常值的特征可作为SARS早期诊断的依据。     

创伤患者血浆干扰素-γ、白介素-4动态变化及其意义

目的　探讨创伤后血浆干扰素 γ(IFN γ)、白介素 4 (IL 4 )的变化规律及其临床意义。方法　创伤患者 35例 ,分别于伤后 2 4小时内、3、5、7天取血 ,采用酶联免疫吸附法 (ELISA)测定血浆白介素 4 (IL 4 )和干扰素 γ(IFN γ)含量 ,并分析IL 4和IFN γ水平以及IFN γ/IL 4比值与患者伤情、严重并发症及预后的关系。结果　创伤患者血中IFN γ含量于伤后第 5天起开始下降 ,至第 7天最明显 ;伤后血中IL 4含量呈现波动性上升 ;IFN γ/IL 4比值呈持续性下降 ,其中MODS组下降较非MODS组显著 (P <0 .0 5 ) ;IFN γ/IL 4比值与APACHEⅡ评分呈负相关 (r=- 0 .6 2 2 ,P <0 .0 5 ) ,与患者感染率呈显著负相关 (r=- 0 .86 8,P <0 .0 1 )。结论　创伤应激后可出现IFN γ/IL 4持续性下降 ,与患者的免疫机能、预后情况密切相关     

放射损伤和放烧复合伤影响小鼠脾脏T细胞IFN-γ和IL-4基因表达的实验研究

目的 阐明机体免疫功能的放烧复合效应，为放烧复合伤免疫功能障碍的调控提供依据。方法 于不同时相点测定放烧复合伤小鼠（5Gy γ射线照射合并12％Ⅱ度烧伤）的脾体比，脾脏组织学观察，分离脾脏T细胞，RT－PCR检测其IFN－γ和IL－4的基因表达水平。结果 ①伤后放烧组的脾脏指数和组织结构的恢复明显快于单放组；②放射损伤后T细胞IFN－γ和IL－4的基因表达均受到抑制，但与IL－4比较，IFN－γ表达量的降幅小、恢复快；③放射损伤后脾脏指数和T细胞IL－4的基因表达呈双向变化；④放烧组IFN－γ表达量明显高于单放组，而两伤组IL－4表达量无区别。结论 ①放射损伤是放烧复合伤的主导损伤因素；②IL－4的辐射敏感性高于IFN－γ；③烧伤因素能促进放烧复合伤后脾脏淋巴细胞的增殖及IFN－γ的表达，不能促进IL－4的表达；④受照机体内仍然存在着促进IL－4表达恢复的有效机理。     

HLA-DRB1*07与广东汉族人慢性乙型肝炎相关性的研究

为探讨HLA DRB1 0 7与广东汉族人慢性乙型肝炎的相关性 ,收集HBV慢性携带者 10 0例、自动清除者 2 1例和非HBV暴露者 70例作为研究对象 ,采用序列特异性引物聚合酶链反应法检测HLA DRB1 0 7,双抗体夹心法检测IFN γ和IL 4。结果发现 ,慢性携带者HLA DRB1 0 7的阳性率为 30 % ,高于自动清除组 (4 76 % ,P <0 0 5 )和非暴露组 (8 5 % ,P <0 0 5 ) ;HBV慢性携带者中 ,HLA DRB1 0 7阳性者外周血IFN γ表达水平为 1132 0 4±75 36pg/ml,IL 4表达水平为 876 79± 4 7 5 3pg/ml,而HLA DRB1 0 7阴性者IFN γ表达水平为 14 32 10±198 13pg/ml,IL 4表达水平为 6 81 99± 6 1 5 9pg/ml,HLA DRB1 0 7阳性者外周血IL 4表达水平高于阴性者 (P <0 0 5 ) ,而二者之间IFN γ的表达水平无显著差异。结果表明HLA DRB1 0 7与广东汉族人慢性乙型肝炎关联     

粒细胞集落刺激因子(G-CSF)对老年急性心肌梗死患者外周血干细胞动员效率的临床观察

目的　观察粒细胞集落刺激因子 (G CSF)对老年 (≥ 70岁 )急性心肌梗死 (AMI)患者 ,外周血干细胞动员效率。方法　 10例老年AMI患者予以G CSF 30 0 μg～ 6 0 0 μg d皮下注射 ,连续 5d ,第 6天经美国Baxter公司生产的CS30 0 0PLUS血细胞分离机 ,分离PBSC ,动员前及动员后第 3～ 7天行外周血WBC计数检查 ,经流式细胞仪测定CD34+ 的细胞数量。结果　在动员前及动员后第 3～ 7天外周血中WBC量分别为 6 .75× 10 9 L、2 8.16× 10 9 L、34.93× 10 9 L、34.4 0× 10 9 L、38.93× 10 9 L、2 1.85× 10 9 L ;CD34+ 量分别为 6 .2 5× 10 6 、5 1.10× 10 6 、92 .6 0× 10 6 、10 9.6 5× 10 6 、134.6 9× 10 6 、4 5 .0 9× 10 6 ;在动员前及动员后第 3～ 7天外周血中WBC、CD34+ 量与动员时间变化曲线均显示曲线高峰在动员后第 6天 ;患者外周血中CD34+ 细胞数量与WBC数量变化呈正相关 (r=0 .94 0 ) .结论　老年AMI患者行G CSF动员安全可行 ,外周血中CD34+ 细胞数量与WBC变化曲线高峰均出现在第 6天 ,且WBC与CD34+ 细胞数量之间具有正相关     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.