应用YAC探针进行荧光原位杂交检测慢性粒细胞白血病BCR基因重排

为检测慢性粒细胞白血病（ＣＭＬ）中ＢＣＲ基因重排，采用酵母人工染色体（ＹＡＣ）ＤＮＡ的Ｉｎ－ｔｅｒ－Ａｌｕ－ＰＣＲ产物为探针进行荧光原位杂交（ＦＩＳＨ）研究了１０例ＣＭＬ，其中包括初诊患者２例，ＣＭＬ急变并接受化疗２例，α干扰素治疗２例，自体骨髓移植术（ＡＢＭＴ）后３例和Ｐｈ染色体阴性ＣＭＬ１例。同时进行细胞遗传学和ＲＴ－ＰＣＲ检测。结果：９例ＣＭＬ的４６％～１００％的可分析核型显示ｔ（９；２２）易位，其中携带ｔ（９；２２）细胞最少者为１例自体骨髓移植术后８个月的患者，其６４％的分裂相存在ｔ（９；２２），３６％为正常核型；１例Ｐｈ（－）ＣＭＬ未见ＢＣＲ基因易位，而ＲＴ－ＰＣＲ（＋），提示ＡＢＬ基因片段插入２２ｑ１１，造成隐匿性Ｐｈ染色体。结果表明：应用ＹＡＣ探针进行原位杂交的定位明确。ＦＩＳＨ检测微小残留病（ＭＲＤ）比常规细胞遗传学方法更敏感，而且可以完成ＰＣＲ方法不易进行的定量分析。     

THE HEMORHEOLOGY CHANGE IN ISCHEMIC REGION BY CORONARY STENOSIS AFTER LASER IRRADIATION ON BLOOD

ＴＨＥＨＥＭＯＲＨＥＯＬＯＧＹＣＨＡＮＧＥＩＮＩＳＣＨＥＭＩＣＲＥＧＩＯＮＢＹＣＯＲＯＮＡＲＹＳＴＥＮＯＳＩＳＡＦＴＥＲＬＡＳＥＲＩＲＲＡＤＩＡＴＩＯＮＯＮＢＬＯＯＤＴＨＥＨＥＭＯＲＨＥＯＬＯＧＹＣＨＡＮＧＥＩＮＩＳＣＨＥＭＩＣＲＥＧＩＯＮＢＹＣＯＲ...     

ELECTRON MICROSCOPE ANALYSIS OF MITOCHONDRIA ON RENAL TUBULE CELL SUFFERED WARM ISCHEMIC AND REPER FUSIVE DAMAGE IN RABBITS

ＥＬＥＣＴＲＯＮＭＩＣＲＯＳＣＯＰＥＡＮＡＬＹＳＩＳＯＦＭＩＴＯＣＨＯＮＤＲＩＡＯＮＲＥＮＡＬＴＵＢＵＬＥＣＥＬＬＳＵＦＦＥＲＥＤＷＡＲＭＩＳＣＨＥＭＩＣＡＮＤＲＥＰＥＲＦＵＳＩＶＥＤＡＭＡＧＥＩＮＲＡＢＢＩＴＳＥＬＥＣＴＲＯＮＭＩＣＲＯＳＣＯＰＥＡ...     

A FREQUENCY DOMAIN QUANTITATIVE MEASUREMENT METHOD FOR BLOOD FLOW VELOCITY WITH BIDIRECTIONAL DOPPLERULTRASOUND AVOIDING DOWN-BAND CHANNEL

ＡＦＲＥＱＵＥＮＣＹＤＯＭＡＩＮＱＵＡＮＴＩＴＡＴＩＶＥＭＥＡＳＵＲＥＭＥＮＴＭＥＴＨＯＤＦＯＲＢＬＯＯＤＦＬＯＷＶＥＬＯＣＩＴＹＷＩＴＨＢＩＤＩＲＥＣＴＩＯＮＡＬＤＯＰＰＬＥＲＵＬＴＲＡＳＯＵＮＤＡＶＯＩＤＩＮＧＤＯＷＮ－ＢＡＮＤＣＨＡＮＮＥＬＷａ...     

MICROPROCESSOR-BASED SYSTEM FOR MEASUREMENT OF BIOELECTRICAL IMPEDANCE DURING HEMODIALYSIS

ＭＩＣＲＯＰＲＯＣＥＳＳＯＲ－ＢＡＳＥＤＳＹＳＴＥＭＦＯＲＭＥＡＳＵＲＥＭＥＮＴＯＦＢＩＯＥＬＥＣＴＲＩＣＡＬＩＭＰＥＤＡＮＣＥＤＵＲＩＮＧＨＥＭＯＤＩＡＬＹＳＩＳＭＩＣＲＯＰＲＯＣＥＳＳＯＲ－ＢＡＳＥＤＳＹＳＴＥＭＦＯＲＭＥＡＳＵＲＥＭＥＮＴＯＦＢ...     

PRELIMINARY RESEARCH OF HUMAN HEPATOCYTE CELL LINE AS THE BIOLOGICAL MATERIAL OF ELAD

ＰＲＥＬＩＭＩＮＡＲＹＲＥＳＥＡＲＣＨＯＦＨＵＭＡＮＨＥＰＡＴＯＣＹＴＥＣＥＬＬＬＩＮＥＡＳＴＨＥＢＩＯＬＯＧＩＣＡＬＭＡＴＥＲＩＡＬＯＦＥＬＡＤＰＲＥＬＩＭＩＮＡＲＹＲＥＳＥＡＲＣＨＯＦＨＵＭＡＮＨＥＰＡＴＯＣＹＴＥＣＥＬＬＬＩＮＥＡＳＴＨＥＢＩＯ...     

EXPERIMENTAL STUDIES ON HIGHDENSITY ENDOHELIAL CULTURES IN ARTIFICIAL BLOOD VESSELS

ＥＸＰＥＲＩＭＥＮＴＡＬＳＴＵＤＩＥＳＯＮＨＩＧＨ－ＤＥＮＳＩＴＹＥＮＤＯＨＥＬＩＡＬＣＵＬＴＵＲＥＳＩＮＡＲＴＩＦＩＣＩＡＬＢＬＯＯＤＶＥＳＳＥＬＳＥＸＰＥＲＩＭＥＮＴＡＬＳＴＵＤＩＥＳＯＮＨＩＧＨ－ＤＥＮＳＩＴＹＥＮＤＯＨＥＬＩＡＬＣＵＬＴＵＲＥ...     

EFFECT OF SUSPENDING MEDIUM VISCOSITY ONORIENTATION AND DEFORMATION OF RBCs IN A SHEAR FlOW FIELD

ＥＦＦＥＣＴＯＦＳＵＣＰＥＮＤＩＮＧＭＥＤＩＵＭＶＩＳＣＯＳＩＴＹＯＮＯＲＩＥＮＴＡＴＩＯＮＡＮＤＤＥＦＯＲＭＡＴＩＯＮＯＦＲＢＣＳＩＮＡＳＨＥＡＲＦｌＯＷＦＩＥＬＤＷｅｎＺｏｎｇ－ｙａｏ，ＭａＷｅｉｙｕａｎ，ＧａｏＴｉｅ，ＳｕｎＤａｇｏｎｇＤｅｐａ...     

BIOLOGIC HEMOSTATIC GLUE IN GENERAL SURGERY

ＢＩＯＬＯＧＩＣＨＥＭＯＳＴＡＴＩＣＧＬＵＥＩＮＧＥＮＥＲＡＬＳＵＲＧＥＲＹＢＩＯＬＯＧＩＣＨＥＭＯＳＴＡＴＩＣＧＬＵＥＩＮＧＥＮＥＲＡＬＳＵＲＧＥＲＹＨｕＹｉ－ｚｅ（ＤｅｐａｒｔｍｅｎｔｏｆＳｕｒｇｅｒｙ，ＳｅｃｏｎｄＡｆｆｉｌｉａｔｅｄＨｏｓｐｉ...     

STUDY ON MECHANISM OF THE BIOEFFECTS OF TRANSIENT ELECTROMAGNETIC PULSE

ＳＴＵＤＹＯＮＭＥＣＨＡＮＩＳＭＯＦＴＨＥＢＩＯＥＦＦＥＣＴＳＯＦＴＲＡＮＳＩＥＮＴＥＬＥＣＴＲＯＭＡＧＮＥＴＩＣＰＵＬＳＥＳＴＵＤＹＯＮＭＥＣＨＡＮＩＳＭＯＦＴＨＥＢＩＯＥＦＦＥＣＴＳＯＦＴＲＡＮＳＩＥＮＴＥＬＥＣＴＲＯＭＡＧＮＥＴＩＣＰＵＬＳＥＷ...     

Loss of heterozygosity of the BRCA1 and FHIT genes in patients with sporadic breast cancer from Southern Brazil

Aim: The evaluation of allelic losses at the FHIT and the BRCA1 genes and at three other loci at the 17q region in a series of 34 sporadic breast cancer cases from Southern Brazil. METHODS: The samples were evaluated for loss of heterozygosity (LOH) at the FHIT and the BRCA1 genes and at three other microsatellite markers at 17q, and the findings were correlated with clinicopathological parameters. RESULTS: The BRCA1 intragenic marker, D17S855, had the highest frequency of LOH, detected in 10 of 24 informative cases, followed by the D17S579 (six of 23 informative cases), D17S806 (five of 21 informative cases), and D17S785 markers (five of 21 informative cases). LOH at the FHIT intragenic marker, D3S1300, was found in six of 25 informative cases. In four of the six cases with LOH of the FHIT gene, there was concomitant loss of the BRCA1 intragenic marker. CONCLUSIONS: The frequency of allelic losses in the FHIT and BRCA1 loci in the Southern Brazilian population is similar to that described in the general population. No correlations were found when the total LOH frequency was compared with tumour size, grade, or presence of axillary lymph node metastasis. Further studies using larger sporadic breast cancer samples and additional markers would be useful to confirm these findings, in addition to establishing more specific associations with clinicopathological parameters in this specific population.     

Patterns of allelic loss at the BRCA1 locus in Arabic women with breast cancer

Loss of heterozygosity (LOH) of BRCA1, a tumor suppressor gene, is one mechanism of genetic inactivation in both sporadic and familial forms of breast cancer. Studies reported in breast cancers from women of Northern European descent have shown LOH in 30-50% of sporadic tumors. Microsatellite instability (MSI) has served as evidence for involvement of DNA repair genes. This study investigates the extent of allelic imbalance at the BRCA1 region in Arabic women with breast cancer. Paired normal and tumor tissue were available for DNA analysis in 13 cases. Results using fluorescent tagged primers to microsatellite markers D17S1323, D17S1325 and D17S855 intragenic to BRCA1 were analyzed using an ABI 310 DNA sequencer. As compared to normal DNA, MSI and LOH were recognized as a gain and a loss, respectively, of one signal in one allele in the tumor DNA. Microsatellite analyses showed 12 of 13 (92%) cases with LOH or MSI or both. Three cases demonstrated LOH alone, 3 cases with MSI alone. Six cases indicated both LOH and MSI; 2 cases with either LOH or MSI in separate markers. The combined finding of LOH and MSI in the same marker was detected only with D17S1325 in 4/6 cases. The proportion of aberrant findings of the BRCA1 locus in breast cancer appears to be higher in Arabic women than in other populations studied to date.     

Abnormalities of chromosome 17 in oesophageal cancer

BACKGROUND: Oesophageal cancer is the most common malignancy encountered in South African males, especially in the Eastern Cape and surrounding region of South Africa. There are a number of risk factors and predisposing conditions that have been implicated in the aetiology of the disease. The tylosis oesophageal cancer (TOC) gene, localised to a small region on chromosome 17q25, has been shown to be associated with oesophageal squamous cell carcinoma. AIM: To investigate loss of heterozygosity (LOH) and microsatellite instability (MSI) in the region of the TOC locus. METHODS: In 74 oesophagectomy specimens for squamous cell carcinoma, microsatellite PCR was performed using five fluorescently labelled TOC markers. The PCR products were analysed and the data correlated with clinicopathological findings. RESULTS: LOH ranged from 25% to 60%. LOH for the individual markers was as follows: D17S1839, 25%; D17S1864, 36%; D17S1817, 38%; D17S785, 47.8%; and D17S579, 60%. MSI ranged from 4.1% to 6.8% for the five loci in the 17q region. MSI was 4.1% for the markers D17S579, D17S785 and D17S1817. Marker D17S1864 showed MSI to occur in 4 cases (5.4%) and marker D17S1839 in 5 cases (6.8%). CONCLUSION: No significant relationship between genetic and clinical parameters was observed; however, aberrations in poorly differentiated tumours were high for markers D17S579 and D17S1864 (25% and 37%, respectively), indicating that these markers may have an underlying role in the molecular pathogenesis of oesophageal squamous cell carcinoma. In addition, 63% of patients who died showed LOH for the markers D17S579, D17S1864 and D17S1817.     

Microsatellite analysis of breast carcinoma and corresponding local recurrences

Local recurrence is a serious complication of breast carcinoma that reduces quality of life and influences prognosis. The aim of this study was to determine whether local recurrences of breast carcinoma are genetically related to the primary tumours. Forty cases of locally recurrent breast carcinomas (median onset: 3.6 years after primary surgery) were analysed: 22 patients had undergone breast-conserving therapy and 18 mastectomy. Eighteen microsatellites on chromosomes 2p, 3p, 5q, 10q, 11p, 11q, 13q, 17q, 17p, 18p were amplified by PCR using fluorescent-labelled primers, automatically detected after polyacrylamide gel electrophoresis and analysed for loss of heterozygosity (LOH) or microsatellite instability (MSI). Follow-up data were available for 39 cases with a median value of 89 months. All LOH and MSI found in the primary tumours were also present in the corresponding recurrences, indicating that they are genetically related to the primary tumours and not secondary malignancies in the same breast. MSI was found in three cases, of which one harboured MSI at more than two loci. The median value of LOH per case was significantly higher in the recurrent (four per case) compared to the primary tumours (two per case; p < 0.001, Mann-Whitney test), reflecting the genotype of tumour progression. Early local recurrence was associated with specific LOH for TP53.15 (p = 0.018, log-rank test) in the primary tumours. LOH on D13S1699 or D17S855 was associated with lymph node metastases (p = 0.024 and p = 0.019, respectively; chi-square test). In addition, tumour grade, lack of oestrogen or progesterone receptor expression, young patient age and early appearance of local recurrence significantly correlated with poor survival. The development of local recurrence despite clear resection margins may result from residual DCIS distant from the invasive carcinoma, homing of circulating tumour cells, or genetically altered, histologically normal breast tissue not immediately adjacent to the invasive carcinoma.     

中国早发性乳腺癌患者中BRCA1基因突变分析

目的研究中国早发性乳腺癌患者中BRCA1基因致病性突变的发生情况以及家族史在突变携带者识别中的作用。方法研究对象为来自中国4个乳腺癌临床医疗中心的188例早发性乳腺癌病例(发病年龄≤40岁),其中39例(20.1%)有乳腺/卵巢癌家族史。从外周静脉血提取基因组DNA,对BRCA1基因的全部编码区和外显子/内含子拼接区进行PCR扩增。其中22例通过单链构象多态方法进行突变初筛,166例用变性高效液相色谱分析进行初筛;对发现的异常片段通过DNA直接测序的方法进行确认。对发现重复出现突变的样本,选取5个与BRCA1基因连锁的标记(D17S855、D17S1322、D17S1323、D17S1326和D17S1327)进行等位基因型分析。结果在15例(8.0%)患者中发现有12个BRCA1基因的致病性突变,其中BRCA11100delAT和5589del8突变分别在3个和2个患者中发现。在39例同时伴有乳腺/卵巢癌家族史的病例中共发现有9例(23.1%)携带突变。有(无)乳腺癌家族史的早发性乳腺癌病例间BRCA1基因的突变率的差异有统计学意义(P=0.001)。重复出现的突变在所有检测病例中出现的频率为2.7%,在所有检测到的突变中占33.3%。两个来自中国北方的BRCA11100delAT突变携带病例有相同的等位基因型,而与来自上海地区的此突变携带者的等位基因型在D17S1322位点上有所差异。两例5589del8突变携带者在所检测的5个STR位点上有完全相同的等位基因型。结论这是到目前为止较大规模的关于中国早发性乳腺癌人群的BRCA1基因突变的研究,有助于增加对中国早发性乳腺癌人群中BRCA1基因致病性突变分布的全面认识。在中国早发性乳腺癌人群中,BRCA1基因致病性突变在肿瘤的发生中有比较重要的意义,尤其在伴有乳腺/卵巢癌家族史病例中该基因突变的意义尤为突出。两个重复出现的突变可能在中国人群中有始祖效应,在进行全基因检测前对其先进行检测可能非常合算。     

Loss of heterozygosity in P53, BRCA1, and estrogen receptor genes and correlation to expression of p53 protein in ovarian epithelial tumors of different cell types and biological behavior

Loss of heterozygosity (LOH) of tumor suppressor genes (TSGs) in ovarian epithelial tumors of differing cell types and biological behavior has not been thoroughly investigated. Moreover, there have been conflicting reports correlating LOH of the p53 gene to overexpression of p53 protein. This study evaluated 34 formalin-fixed, paraffin-embedded ovarian epithelial tumors for LOH by polymerase chain reaction (PCR) for the following microsatellite markers: TP53(17p13.1/p53 gene), D17S579(17q/BRCA1 gene), and ESR (6q24-27/estrogen receptor gene). LOH of the TP53 marker was detected in 4 (44%) of 9 informative serous cystadenocarcinomas (SCa) but in 0 of 4 informative clear cell carcinomas (CCa) and 0 of 5 informative serous tumors of low malignant potential (SLMP). LOH of the BRCA1 marker was detected in 5 (83%) of 6 informative SCa, but in 1 (13%) of 8 informative CCa and 1 (14%) of 7 informative SLMP. LOH of the ESR marker was detected in 4 (50%) of 8 informative SCa, but in 0 of 4 informative CCa and 1 (16%) of 6 informative SLMP. p53 protein overexpression was present in 8 of 12 SCa but did not correlate to TP53 LOH. LOH for TP53, D17S579/ BRCA1, and ESR is common in ovarian SCa, and is observed in primary tumors as well as metastases. In contrast, these genetic alterations are less common in CCa and in the biologically less aggressive SLMP tumors. These data suggest different mechanisms of oncogenesis in ovarian epithelial tumors of different cell types and biological behavior.     

Frequent microsatellite alterations in a predominantly younger age of onset breast cancer population

Microsatellite instability (MSI) and loss of heterozygosity (LOH) was investigated in paired tumour and normal tissue DNA from 108 predominantly premenopausal breast cancer patients (under age 45 years at presentation) for 25 simple repeat loci interspersed across 11 chromosomes. MSI was observed at a single locus in 69 (64%) patients; 41 of these had instability at more than one site. Greatest frequency of MSI was at loci D2S1356 (33%), D2S2739 (22%), D3S1766 (21%) and D13S796 (20%). LOH was seen at a single site in 55% of patients and at two or more sites in 27 patients with greatest frequency at D2S1356 (33%), D2S443 (19%) and D17S1299 (18%). Both mutations were found in the same patient but at different loci. Clearly, choice of loci is a determining factor in assessing genomic instability. The relatively high frequency of MSI may also reflect peculiarities of this younger patient population. Occurrence of MSI or LOH was unrelated to clinical stage, nodal status, tumour size or grade or steroid receptor status. It was independent of mutations detected in exons 5-9 of the p53 gene. There was no significant association with survival. The lack of such correlations reflects a random disabling mechanism that may equally affect genes promoting cell death as well as growth.     

Loss of heterozygosity of p53, BRCA1, VHL,and estrogen receptor genes in breast carcinoma: correlation with related protein products and morphologic features

Correlation of loss of heterozygosity (LOH) of p53, BRCA1, VHL, and estrogen receptor (ER) genes with the expression of related protein products and morphologic features predictive of aggressive biologic behavior was investigated to determine the significance of LOH in these genes. DNA from 35 formalin-fixed, paraffin-embedded breast carcinomas was obtained by microdissection of histologic sections. LOH was determined by polymerase chain reaction (PCR) using primers TP53, D3S1038, D17S855, and ESR for p53, VHL, BRCA1, and ER genes, respectively. p53, ER, and progesterone receptor (PR) protein expression was evaluated by immunohistochemistry. Morphologic evaluation included histologic type, and histologic and nuclear grades. TP53 LOH was identified in 13 (52%), BRCA1 LOH in 3 (17%), VHL LOH in 1 (4%), and ER LOH in 4 (21%) of 25, 17, 24, and 19 informative cases, respectively. p53 and ER protein expression was identified in 20 (57%) and 25 (71%) cases, respectively. TP53 LOH directly correlated with both high histologic and nuclear grade (P<0.01). BRCA1, VHL, and ER LOH was not frequent enough for correlation to morphologic features. Although 4 of 4 ER and 7 of 13 p53 LOH cases expressed related proteins, LOH did not correlate with protein expression. TP53 LOH may be an event contributing to aggressive biologic behavior since it is strongly associated with high histologic and nuclear grade. Missense or nonsense mutations may explain the absence of detectable p53 protein in 6 of 13 cases with p53 LOH. All 4 ER LOH cases expressed ER protein. BRCA1 and VHL LOH is infrequent in sporadic breast carcinoma.     

German family study on hereditary breast and/or ovarian cancer: Germline mutation analysis of the BRCA1 gene

Women harboring BRCA1 germline mutations carry an 85% lifetime risk of developing breast cancer and a 63% risk of ovarian cancer. In this first systematic study of familial breast and/or ovarian cancer in Germany we investigated 29 families for germline mutations in the BRCA1 gene. We identified mutations in three breast cancer families and in four breast-ovarian cancer families. The mutations include one missense mutation, one frameshift mutation, one splice mutation, and four nonsense mutations cosegregating with breast and/or ovarian susceptibility in five of ten (50%) families showing positive evidence of linkage to chromosome band 17q21 and in two of 19 (11%) families where linkage data was not available. Two apparently unrelated families carried the same nonsense mutation at codon 1835 and three families harbored a C to T transition at nucleotide 49 of the untranslated exon 4. Allelotyping of the markers D17S855, D17S1322, D17S1323, and D17S1327 located within or near BRCA1 revealed that all affected individuals in the two families harboring the mutation at codon 1835 shared at least one allele indicating a founder mutation. With respect to the overall mutation spectrum, no mutations were identified in exon 11 (0/7) in this set of German families. These findings differed significant from those in British (17/32)(P = 0.012) and Southern Swedish (13/15) (P < 0.001) families. The lack of BRCA1 mutations in exon 11 which represents 61% of the entire coding sequence may provide additional insight into BRCA1 associated breast and ovarian tumor development. Genes Chromosom. Cancer 18:126–132, 1997. © 1997 Wiley-Liss, Inc.     

Microsatellite instability in ovarian and other pelvic carcinomas

Twenty-six cases of ovarian carcinoma and six cases of other pelvic neoplasms were analyzed for microsatellite instability (MSI) using frozen specimens, fluorescence technology, and four selected markers (D2S123 on chromosome 2, D18S58 on chromosome 18, BAT26 on chromosome 2, and BAT40 on chromosome 1). This procedure also allowed the detection of loss of heterogeneity (LOH) at the four selected loci. One of the cases of ovarian carcinoma exhibited MSI and this was evident at three loci. Of 44 informative loci, 7 exhibited LOH representing 3 cases of ovarian carcinoma, 3 of 4 cases of primary peritoneal carcinoma, and one case of unknown primary. These data support other findings that MSI is not a frequent occurrence in ovarian cancer; however, LOH is a more frequent event and may be a target for the development of diagnostic/prognostic procedures for ovarian and primary peritoneal carcinoma.