A Comparative Study on the Effect of BCG-PSN and Thymopeptides on T-lymphocyte Subsets of Normal and Immunosuppressed Mice

Polysaccharidenucleicacidfractionofbacilluscalmetteguerin (BCG PSN ,SiqikangInjection)andthymopeptidesarenowtwowidelyusedimmunomod ulatorsinclinicalpractice .Theyareusuallyusedasanadjuvanttherapyforvirusinfection ,autoimmunediseasesandneoplasms ,whichhavebeenclinicallyprovedtobeeffective .Somereportsdemonstratedthattheybothcanstimulatetheproliferationanddif ferentiationofT lymphocytes.However ,theexactmechanismshavenotbeenelucidatedyet .InordertocomparetheirmodulatingmechanismsonT lympho c…     

特发性脊柱侧凸King型和型远端融合椎的选择

目的回顾采用椎弓根钩和螺钉CD技术治疗特发性脊柱侧凸King型和型患者,分析选择性缩短融合节段的治疗效果。方法2000年3月~2003年1月,治疗58例特发性脊柱侧凸单胸弯患者,男17例,女41例,年龄12~18岁,平均14岁。其中King型40例,King型18例。胸弯Cobb角平均64°(50~83°),柔韧性62%;腰弯Cobb角平均37°(16~48°),柔韧性105%。腰弯腰骶角平均为17°(10~22°)。所有患者C7重力垂线均不同程度地偏离骶骨中线。采用椎弓根钩和螺钉CD技术矫形治疗,以中立椎为基础选择远端融合椎,所有远端融合节段均未超过中立椎。术后随访摄站立前后位和侧位X线片,观察各项指标的变化。结果患者均获随访1年8个月~3年2个月,平均2.4年,均未出现明显的躯干侧方移位和双肩不平衡。术后Cobb角平均丢失3.1°(-1~5°);最后随访时,胸弯矫正率68%;除2例C7重力垂线偏离骶骨中线1~2cm外,其余均通过骶骨中线;腰弯腰骶角减少至平均8°(2~13°),矫正率为53%;48例远端融合椎为非稳定椎者术后成为稳定椎。与Harrington远端融合椎选择原则相比,患者远端融合椎平均节省1.4个节段(1~2个节段)。结论采用三维节段性器械内固定系统治疗特发性单胸弯时,以中立椎为基础选择远端融合椎,可获得较好的临床效果。     

Novel multi-probe RNase protection assay set for detection of endotoxin associated receptors gene expression

Objective: To construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. Methods : The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR)using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7/ promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method. Results: The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation. Conclusions: These new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.     

乳凝集素对新生仔兔小肠粘膜及小肠CD4~+,CD8~+,CD25~+淋巴细胞发育的影响

目的探讨乳凝集素(lactadherin)对新生仔兔在吮乳期及断奶期小肠粘膜发育以及小肠淋巴细胞发育的影响。方法新生仔兔共45只,随机分为母乳组(GB),配方奶组(GF)及配方奶加乳凝集素组(GL,100μg/ml),每组15只,在3w和5w应用图像分析技术检测新生仔兔小肠粘膜发育情况,分别在1、2、3、4和5w时,应用流式细胞技术测定小肠CD4+,CD8+,CD25+淋巴细胞发育情况。结果(1)GL组3w空肠绒毛周长、绒毛面积、绒毛高度及隐窝深度均超过GF组,GB组3w空肠绒毛周长及绒毛高度超过GF组;GL组5w空肠和回肠绒毛周长、绒毛面积、绒毛高度及隐窝深度均超过GF组和GB组。(2)5w各组小肠绒毛发生变化,绒毛表面由高密度的手指状变为平滑叶状或舌状,GL组绒毛面积和高度恢复较GB和GF组好。(3)GL组回肠CD4+淋巴细胞发育较GF组好,GB组较GL组好;GL组回肠CD25+淋巴细胞发育较GF组好(P<0.05)。结论乳凝集素对近端空肠粘膜形态发育和小肠CD4+和CD25+淋巴细胞发育有促进作用,并且可能帮助断奶期绒毛的恢复。     

CD4+ lymphocyte subsets in the cerebrospinal fluid of multiple sclerosis and non-inflammatory neurological diseases

Summary We studied paired cerebrospinal fluid (CSF) and peripheral blood (PB) samples from 18 inactive multiple sclerosis (MS) patients and 10 with non-inflammatory neurological diseases. By means of a dual-colour cytofluorimetric micromethod we were able to count 1500 cells on average in each CSF sample. We found a significant reduction of CD45RA+ and CD4+CD45RA+ cells in the CSF of MS patients. Similarly, CD45RA+ and CD4+CD45RA+ CSF/PB ratios were lower compared with controls. The reduction of suppressor-inducer T-cells did not correlate with CD8+ cell levels in the CSF. The CD4+ subset ratio (CD4+CD45RA–/CD4+CD45RA+) was significantly increased in the CSF of MS patients. Our data suggest that the reduction of CD4+CD45RA+ cells in the PB is not due to a segregation of such cells in the CSF. Conversely, CSF changes reflect changes in the PB similar to these found for other T-cell subsets.     

协同刺激分子CD28在结核杆菌抗原体外激活人外周血γδT细胞中的作用

目的 :为了探讨CD2 8协同刺激分子在结核杆菌 (Mtb)低分子多肽抗原体外激活人外周血γδ T细胞中的作用。方法 :采用激发型抗CD2 8单抗模拟第二信号 ,Mtb低分子多肽抗原作为刺激原 ,对纯化的人外周血T细胞进行体外刺激和培养 ;用流式细胞仪检测γδ T细胞上CD2 8分子的表达、γδ T细胞的增殖效应及活化的γδ T细胞上CD6 9分子的表达。结果 :人外周血γδ T细胞中有 5 0 %左右表达CD2 8分子 ;抗CD2 8单抗协同Mtb抗原可刺激γδ T细胞的活化和增殖 ;但抗CD2 8单抗或Mtb抗原单独刺激则无作用。活化的γδ T细胞表面表达CD6 9分子。结论 :Mtb抗原在选择性活化人外周血γδ T细胞时需要第二信号的参与 ;CD2 8在Mtb抗原激活γδ T细胞时可提供协同刺激信号 ;CD6 9可作为γδ T细胞的早期活化标志。     

重组跨膜型人CD55分子在小鼠NIH3T3细胞上的表达及其补体溶破保护功能的研究

目的 :在小鼠NIH3T3细胞转染表达人天然GPI锚固型CD5 5和重组跨膜型CD5 5 TM分子 ,观察比较它们对人补体溶破异源细胞的抑制功能。方法 :将带有CD5 5cDNA、CD5 5 TMcDNA的重组逆病毒表达质粒CD5 5 pLXSN、CD5 5TM pLXSN经脂质体法转染PA317细胞 ,用病毒上清感染小鼠成纤维母细胞NIH3T3。经G418加压筛选 ,利用FACS检测获得表达CD5 5和CD5 5 TM分子的阳性细胞克隆 ,通过MTT比色法比较两种分子对人血清补体溶破细胞的抑制功能有无差别。结果 :细胞转染筛选获得多个表达跨膜型人CD5 5分子的NIH3T3细胞克隆 ,补体杀伤试验证实其具有抑制人补体溶破的功能 ,且两种分子的补体抑制功能无明显差异。结论 :成功地建立了稳定表达天然CD5 5、跨膜型CD5 5分子的小鼠NIH3T3细胞 ,证实其表达的GPI型CD5 5分子和CD5 5TM分子均具有抑制人补体溶破细胞的功能 ,为进一步探讨应用跨膜型的CD5 5分子对PNH进行基因治疗奠定了基础。     

共刺激分子CD80和CD86在蠕虫保护性免疫及其Th2免疫应答中作用机制的研究

目的:探讨共刺激分子CD80和CD86在蠕虫保护性免疫及其Th2免疫应答中的作用机制。方法:采用巴西日圆线虫第3期幼虫感染BALB/c小鼠,在感染当天、感染后第3、7d分别腹腔注射大鼠抗CD80和/或抗CD86单克隆抗体,以阻断协同刺激信号。在感染当天,感染后第7、14d,计数小鼠外周血液的嗜酸性粒细胞,于感染后第14d计数小肠绒毛嗜酸性粒细胞;采用WatanabeN法测定IgE。结果:联合应用抗CD80和抗CD86两种单克隆抗体可导致成虫产卵量明显增加,成虫数量显著增多及排虫时间延迟。同时两种单克隆抗体也完全抑制了外周血液及小肠组织的嗜酸性粒细胞的增多并部分抑制了IgE水平的升高。而单独应用抗CD80或抗CD86McAb对保护性免疫及Th2型免疫应答反应均无明显影响。结论:在蠕虫保护性免疫和Th2型免疫应答反应中,T细胞的活化需要CD80和CD86两种共刺激分子的参与;而CD80或CD86单一刺激分子即可提供足够的共刺激信号。研究提示,人为调控CD80和CD86共刺激分子可能有助于对蠕虫病及Th2型免疫应答所介导的其它疾病的治疗。     

三七多糖对创伤大鼠CD+4/CD+8和IL-2水平的影响(摘要)

观察三七多糖对创伤大鼠免疫功能的影响.方法:40只SD大鼠,随机分为正常对照组(A)、创伤对照组(B)、创伤后三七多糖28.5 mg·kg-1.     

Expression of CD44V2 in transitional cell carcinoma of the urinary bladder and in urine

CD44 is the principal cell surface receptor for hyaluronate. Variant forms of the receptor, produced by alternative splicing, have been found to be associated with tumor progression in a variety of cancers. Based on investigations at the RNA level, it has recently been proposed that expression of CD44 variant V2 was present in urothelial cancer but not in normal urothelium. Since a distinctive marker for urothelial cancer would be extremely useful, frozen sections of normal urothelium and urothelial cancer were examined for expression of standard CD44 and CD44V2. Frozen sections of specimens of 35 patients with transitional cell carcinoma of the bladder, 16 specimens of normal bladder and 5 ureters were examined. Immunohistochemical staining was performed using a polyclonal antibody to CD44V2 (PAB CD44V2), a monoclonal antibody to CD44V2 (MAB CD44V2) and a monoclonal antibody to CD44S (MAB CD44S). CD44V2 and CD44S were also measured in lysates of urine sediments from 21 patients by enzyme-linked immunoabsorbent assay (ELISA). All investigated transitional cell carcinomas expressed CD44V2. There was no differentiation between invasive and noninvasive carcinoma. CD44V2 was also expressed in normal urothelium. Standard CD44 was expressed by the transitional cell carcinoma, normal urothelium, musculature and interstitial tissue. The amount of CD44V2 and CD44S in lysates of urine sediments is not correlated to diagnosis. In contrast to investigations at the RNA level, CD44V2 on the protein level seems not to be a distinctive marker for urothelial cancer. Therefore, CD44V2 will not be a useful diagnostic marker for detection of transitional cell carcinoma.