急性白血病非系限性分化抗原的动态研究

为探讨仅表达非系限性分化抗原的急性白血病的细胞起源，采用单克隆抗体免疫酶标和聚合酶链反应技术分析了１２例初诊时仅表达非系限性分化抗原的急性白血病化疗后免疫表型及免疫球蛋白重链（ＩｇＨ）和Ｔ细胞受体（ＴＣＲ）γ基因重排的变化。结果表明：初诊时仅表达ＣＤ３８抗原的５例急性白血病，化疗后３例出现Ｔ细胞相关抗原表达，并伴ＴＣＲγ基因重排；５例初诊时仅表达ＨＬＡ－ＤＲ或ＣＤ９的急性白血病，化疗后４例出现Ｂ细胞相关抗原表达，均伴有ＩｇＨ基因重排；２例无任何抗原表达者，化疗后１例表达Ｂ细胞相关抗原，另１例表达骨髓细胞相关抗原。提示免疫标志的动态研究，有助于初诊时免疫学无法分类急性白血病细胞起源的确定。     

κ及λ轻链测定在多发性骨髓瘤诊断中的应用

目的：寻求准确，可靠的免疫球蛋白及κ和λ轻链测定方法，提高多发性骨髓瘤分型和鉴别诊断水平。     

荧光定量聚合酶链反应在尖锐湿疣诊断中的应用

目的　探讨荧光定量聚合酶链反应 (FQ - PCR)对尖锐湿疣 (CA )的诊断价值。方法　 FQ- PCR检测病理确诊的 CA患者 36份标本和健康人 84份标本 ;FQ- PCR及病理诊断同期检测临床送检病例 2 73份标本 ,并做诊断性试验评价。结果　 36例病理确诊的 CA患者 HPV6和 HPV11的 DNA FQ- PCR全部阳性 ,平均拷贝数为 1.0× 10 7± 1.0× 10 2 / m l;84例健康人 FQ- PCR全部阴性 ,平均拷贝数为 3× 10± 2× 10 / ml。 FQ- PCR与病理诊断符合率为 10 0 % ,CA患者与健康人拷贝数的差异有显著性 (P<0 .0 5 )。FQ- PCR与病理诊断对比得出 :灵敏度为 10 0 % ,特异度为 92 % ,误诊率为 0 .0 8,漏诊率为 0 ,准确度为 98.9% ,阳性预测值为 98.8% ,阴性预测值为 10 0 % ,阳性似然比为 12 .5 ,阴性似然比为 0。结论　 FQ - PCR能准确定量 ,灵敏度高 ,特异性强 ,快速 ,简便 ,可作为尖锐湿疣早期诊断的指标     

Using the polymerase chain reaction to maintain DNA probe inventories in clinical and diagnostic laboratories

Clinical and diagnostic DNA laboratories must maintain a large inventory of DNA probes for use in hybridization studies. The preparation of plasmid DNA and isolation of DNA fragments for use as probes in both expensive and time consuming. We present here a rapid and relatively inexpensive method of producing large amounts of DNA fragments from stocks, using the polymerase chain reaction (PCR). Our experience over the past year using this technique has been very positive and we believe many laboratories could benefit by employing such a labor-saving approach to maintaining DNA probes. The technique uses the bacteriophage M13 DNA sequencing primers to amplify cloned inserts contained in commonly used plasmid vectors. As examples, we illustrate the use of DNA produced in this manner as probes for linkage analysis of the fragile X syndrome and for detection of deletions in the Duchenne muscular dystrophy gene. We have also found that at least two probes can be amplified in the same PCR reaction, allowing the detection of two different restriction fragment length polymorphisms (RFLP) simultaneously. It should be possible for laboratories to devise strategies particular to their individual needs using more than one DNA probe produced in the same PCR reaction to detect RFLP's. Such strategies would need only to consider that the predicted alleles of the multiple polymorphisms do not migrate to the same position during electrophoresis. Stocks of single or multiple probes produced by the PCR could then be maintained for more rapid Southern analyses.     

Stereotactic brain biopsy in AIDS

Summary In the hope of finding a treatable condition, the need for rapid diagnosis in HIV-seropositive patients with brain lesions is apparent. In order to evaluate the efficacy of stereotactic brain biopsy in AIDS patients, we retrospectively studied 25 HIV-infected patients undergoing stereotactic biopsy. Brain lesions were identified with gadolinium-enhanced MRI and/or contrastCT. Brain biopsy was performed using the system of Riechert. From 8 up to 15 small tissue samples from one or two targets were obtained in every patient. The biopsy material was examined cytologically, histologically (including electron microscopy), immunohistochemically and, in part, by animal test and polymerase chain reaction (PCR). A definite diagnosis was achieved in 92%. Diagnosis included primary central nervous system lymphoma (PCNSL) (10), toxoplasmosis (10), progressive multifocal leukoencephalopathy (2) and one case of co-existing toxoplasmosis and cytomegalovirus infection. Two biopsies were non-diagnostic. All PCNSLs showed polymorphic B-cell populations of high malignancy; accurate classification according to the Kiel classification was not possible. In 3 lymphomas Epstein-Barr nuclear antigen (EBNA) 2-mRNA could be detected by PCR and confirmed immunohistochemically by EBNA 2 expression. In 6 cases autopsy confirmed the biopsy diagnosis. Conventional histology was not sufficiently decisive for toxoplasmosis and progressive multifocal leukoencephalopathy, so that immunohistochemistry and animal tests became very important for a final diagnosis. With the help of different morphological and molecular biological techniques stereotactic brain biopsy appears to be an effective method in the diagnosis of HIV-associated brain lesions. In view of the marked radio- and chemosensitivity of PCNSLs it is mandatory to establish an early and accurate histological diagnosis for adequate treatment.     

聚合酶链反应（PCR）对单纯疱疹病毒性脑炎的临床诊断价值

应用聚合酶链反应（ＰＣＲ）检测技术对１１８例颅内感染性疾病患者及３７例无神神经系统疾病患者脑脊液（ＣＳＦ）中的单纯疱疹病毒ＤＮＡ（ＨＳＶ－ＤＮＡ）进行了检测及分型。结果提示：“散发性脑炎”组的阳性率为３８．４６％（２０／５２），细菌、真菌性脑膜炎、其它病毒性脑炎组以及无神经系统疾病组均为阴性。作者认为：①本检测是目前单纯疹病毒性脑炎（ＨＳＥ）较为简便而准确的早期诊断方法之一；②ＨＳＶ分型检测，对病原诊断更具有全面性；③两型单纯疱疹病毒均可引起ＨＳＥ。     

Biochemical analysis of plasma-soluble invariant chains and their complex formation with soluble HLA-DR

The invariant chain (CD74) is preferentially localized in the cytoplasm and regulates the loading of exogenous derived peptides into HLA class II heterodimers. In addition, a small proportion of CD74:class II complexes is also expressed on the cell surface. We identified and quantified soluble CD74 (sCD74) molecules in the plasma and sCD74:sHLA-DR complexes by ELISA. EDTA plasma samples from 86 healthy probands were analyzed. sCD74 could be detected in all samples with a mean concentration of 1.14 relative units±1.04 SD (range 0.17-4.31). Approximately 10% of the samples had increased amounts of sCD74 (3.0 relative units). Complexes of sCD74 and sHLA-DR were detected in all samples and their quantities were positively correlated (r=0.83, p0.001) with the sCD74 concentrations. SDS-PAGE analysis of plasma samples with high sCD74 concentrations (3.0 relative units) revealed four isoforms of sCD74 with molecular weights of 45, 43, 35, 31 kDa corresponding to known sizes of intracellular CD74. However, only molecular weights of the 45 and 43 kDa isoforms of sCD74 are found complexed with sHLA-DR. Our data demonstrate, that CD74 molecules are present in their soluble form in the plasma of healthy probands and form complexes with soluble HLA-DR molecules.     

宫颈癌组织中HPV16 E7序列多态性分析

目的探讨广东地区宫颈癌组织中HPV16肿瘤相关性抗原E7基因序列的多态性.方法采用通用引物PCR直接测序法对宫颈癌标本中的HPV分型,从含有HPV16型的标本中采用自行设计的多重引物通过巢式PCR扩增出HPV16E7,经DNA序列测定法检测其基因变异,进而寻找其热点突变.结果50例宫颈癌组织HPV-DNA的检出率为78%,其中HPV16和HPV18型混合感染18例,单纯HPV16型感染15例.33例含有HPV16型的标本中扩增出25例HPV16E7,其中17例647位核苷酸“T”变异“C”,导致相应的蛋白质由天冬氨酸变异为丝氨酸.结论广东地区宫颈癌组织中HPV16E7DNA序列发生碱基替换的区域主要在647位至846位,热点突变点为Nt647和Nt846.     

API2对黏膜相关淋巴瘤凋亡及凋亡相关蛋白的影响

目的：了解黏膜相关淋巴瘤的凋亡水平及其所涉及到的信号传导通路。方法：收集33例胃肠黏膜相关淋巴样组织(MALT)淋巴瘤病例，通过TUNEL技术原位检测MALT淋巴瘤肿瘤细胞的凋亡水平，通过逆转录聚合酶链反应(RT-PCR)和免疫组织化学染色检测肿瘤的凋亡抑制蛋白2编码基因(API2)和Caspase3 mRNA及蛋白，对API2和Caspase3水平作半定量和定量分析。根据凋亡检测结果将全部病例分为两组，即凋亡细胞数每50个高倍视野小于或等于2的病例组和大于2的病例组，比较2组的API2和Caspase3 mRNA及蛋白。结果：半数以上的MALT淋巴瘤中存在凋亡抑制；凋亡抑制的MALT淋巴瘤病例中API2 mRNA及蛋白水平明显高于没有明显凋亡抑制的病例，但Caspase3 mRNA及蛋白水平在两组病例中表达没有明显的差异。结论：MALT淋巴瘤中存在明显的凋亡抑制，其发生与凋亡抑制因子API2的表达上调有关，但API2对凋亡的调节与Caspase3有关的信号传导通路无明显相关。