腹腔镜子宫切除术的临床研究

目的 探讨腹腔镜筋膜内子宫切除术(CISH)和腹腔镜辅助子宫切除术(LAVH)的临床价值。方法 对32例因各种良性妇科疾病而需行子宫切除术的患者采用CISH术18例，LhVH术14例。对照组30例采用腹式子宫切除术(AH)。结果 与AH术相比，CISH术和LAVH术具有术中出血少，术后镇痛用药少，术后病率少，住院时间短。结论 CISH术和LAVH术由于创伤小，恢复快等优点，值得临床推广应用。     

Chromogenic in situ hybridization for Her‐2/neu‐oncogene in breast cancer: comparison of a new dual‐colour chromogenic in situ hybridization with immunohistochemistry and fluorescence in situ hybridization

Aims: Her‐2/neu testing is used as a marker for Herceptin^® therapy. The aim was to investigate new dual‐colour chromogenic in situ hybridization (CISH), in a large number of breast carcinomas (n = 205) with DNA‐specific dual‐colour probes (ZytoVision, Bremerhaven, Germany) and to compare the results with immunohistochemistry (n = 205) and fluorescence in situ hybridization (FISH) (n = 129). Methods and results: Paraffin‐embedded tissue of 205 patients was used. After immunohistochemistry with a focus on immunohistochemically uncertain cases, Her‐2/neu amplification using dual‐colour CISH (ZytoVision^®) was analysed. Validation by FISH was performed. The results were: immunohistochemistry, 27.8% with strong expression, 53.7% with uncertain overexpression and 18.5% with no expression; FISH, 25.6% amplified and 74.4% negative; CISH, 35.6% amplified, 62.9% negative and 1.5% not evaluable. Comparison of immunohistochemistry with CISH: CISH negative in 100% with immunohistochemistry 0/1+, amplified in 82.5% with immunohistochemistry 3+; 5.9% contradictory results: 4.4% immunohistochemistry 3+ and negative by CISH, 1.5% negative in immunohistochemistry but amplified by CISH; FISH (129 cases), 8.5% contradictory results to immunohistochemistry, 6.2% immunohistochemistry 3+ and negative by FISH, 2.3% negative by immunohistochemistry and amplified by FISH; comparison of CISH and FISH, 94.6% same results, 3.9% different ones, 1.6% CISH not analysable. Conclusions: CISH, using dual‐colour probes (ZytoVision^®) is as good as FISH for Her‐2/neu analysis. The few discrepant results are likely to be caused by polysomy or tumour heterogeneity.     

3种子宫切除术式的临床应用效果比较

目的:探讨开腹子宫切除术(TAH)、腹腔镜下筋膜内子宫切除术(CISH)、腹腔镜辅助阴式子宫切除术(LAVH)3种子宫切除方式的临床应用效果.方法:102例子宫良性病变患者,子宫肌瘤72例,子宫腺肌病30例,分为3组:CISH组32例,LAVH组30例,TAH组40例,对3组患者的手术时间、术中出血量及术后情况进行比较.结果:LAVH、CISH组手术时间较TAH组明显延长,术中出血较TAH组明显减少,术后抗生素使用时间、下床活动时间、肛门排气时间、平均住院日较TAH组明显缩短,差异均有统计学意义(P＜0.05).结论:CISH和LAVH较TAH具有损伤少、恢复快的特点,是有效的子宫切除方式.     

Metastatic Breast Cancer Survival according to HER2 and Topo2a Gene Status

The aim of this study was to determine the relationship between amplification of HER2 (Human epidermal growth factor receptor 2) and Topo2a (topoisomerase 2a) and their influence on prognosis in metastatic breast cancer (MBC) patients. Amplification of both HER2 and Topo2a genes was determined by chromogenic in situ hybridization (CISH) in primary tumor tissue of 71 MBC patients. Starting point for follow-up was the time of diagnosis of metastatic disease. Although there was significant correlation between HER2 amplification and Topo2a alterations, Topo2a amplification was not strictly related to HER2 amplification. Follow-up of patients showed that there was no difference in MBC survival between HER2-nonamplified and HER2-amplified patients for subgroup as whole, but there was significant difference in MBC survival between patients with and without Topo2a amplification. HER2 amplification showed prognostic value in subgroups of patients, as well as Topo2a. Combination of these two genes with different status (nonamplified, amplified, coamplified) indicated that they might have additive effect. Also, it has been shown that Topo2a-amplified cases have poorer survival than Topo2a-nonamplified, when treated with CMF therapy.Topo2a amplification seems to be more promising biomarker of MBC survival, than HER2, and potential marker of resistance to CMF therapy.     

High-risk human papillomavirus infection and p16INK4a protein expression in laryngeal lesions

A total of 88 samples of laryngeal lesions (23 vocal cord nodules (VCNs), 23 papillomas (PAs), 18 dysplasias (DYs), and 24 carcinomas (CAs)) were analyzed for p16INK4a protein (p16) expression by immunohistochemistry and for high-risk human papillomavirus (HR-HPV) infection using chromogene in situ hybridization (CISH) and polymerase chain reaction (PCR). The series comprised 62 males and 26 females, aged 1-87 years (median 55 years). p16 expression was detected in 2 of 23 (9%) VCNs, 18 of 23 (78%) PAs, 9 of 18 (50%) DYs, and 14 of 24 (58%) CAs. Using CISH, HR-HPV DNA was detected in 3 of 23 (13%) VCNs, in 19 of 23 (83%) PAs, in 12 of 18 (67%) DYs, and in 14 of 24 (58%) CAs. HR-HPV DNA was found in six of nine (67%) PAs by PCR. A statistically significant difference in p16 expression and HR-HPV DNA presence detected by CISH was observed between VCNs and PAs (p<0.000001). The sensitivity and specificity of p16 expression for HR-HPV DNA presence detected by CISH was 0.612 and 0.773, respectively. Our study confirms a potential role of HR-HPV infection not only in the pathogenesis of malignant, but also in benign laryngeal lesions.     

Comparison of chromogenic in situ hybridisation with fluorescence in situ hybridisation and immunohistochemistry for the assessment of her-2/neu oncogene in archival material of breast carcinoma

The successful treatment of breast cancer is dependent upon a number of complex factors. Her-2/neu gene amplification is known to be one of the most common genetic alterations associated with breast cancer and its accurate determination has become necessary for the selection of patients for trastuzumab therapy.The aim of this study was to prove the consistency of chromogenic in situ hybridisation (CISH) technique after analyzing the overexpression of the Her-2/neu proto-oncogene in 100 invasive breast carcinomas and by comparing CISH results with immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH). Moreover, it was done to evaluate the possible correlation of estrogen (ERs) and progesterone receptors (PRs), the proliferation marker Ki67 and the tumour suppressor gene p53 with HER-2/neu status of these breast carcinomas.Of the 100 breast carcinomas that were analysed, 22 cases showed HER-2/neu amplification, 66 cases showed no amplification, whereas 12 cases were non-interpretable in both assays (FISH and CISH). Consequently, the overall concordance between FISH and CISH was 100%. Additionally, it was observed that when HER-2/neu gene was overexpressed, there was an association with negative PRs and ERs status, negative p53 protein expression and high Ki67 labelling index.It is concluded that patients with tumours scoring 2+ with the CBE356 antibody (borderline immunohistochemistry-tested cases) would also benefit from CISH as it is shown to be highly accurate, practical and can be easily integrated into routine testing in any histopathology laboratory. Finally, CISH represents an important addition to the HER2 testing algorithm.     

Trisomy 6 in Merkel cell carcinoma: a recurrent chromosomal aberration

We retrospectively investigated 17 cases of primary and metastasizing Merkel cell carcinomas (MCC) from 14 patients using chromosomal in-situ hybridization (CISH) to study the occurrence of trisomy 6 in these lesions. METHODS AND RESULTS: Histological diagnosis on all tumour samples was obtained on haematoxylin and eosin stained sections. Immunohistochemistry was performed with antibodies against pancytokeratin (CAM 5.2), cytokeratin 20 (CK20), MIC2 antigen (CD99), neuron-specific enolase (NSE), and chromogranin A (chrA). Sections (4 microm) of the paraffin-embedded tumours were analysed with alpha-satellite centromeric probes for chromosome 6 or 17 using CISH. The signal was amplified by the Tyramide Signal Amplification (TSA) assay. Immunohistochemically, the tumours showed the same general epithelial neuro-endocrine pattern: 11/13 expressed cytokeratin 20, and 47% exhibited trisomy 6, with no significant difference between primary and metastatic lesions. Incomplete follow-up data did not allow us to establish a prognostic value of trisomy 6, however, this aberration might be an additional diagnostic tool in distinguishing MCC from other small round blue cell tumours. CONCLUSIONS: CISH seems to be a promising adjunctive method to diagnose Merkel cell carcinoma. Trisomy 6 should be investigated more closely in these cases, as has been done for chromosomes 1 and 11. Of particular interest would be identification of modifications in proto-oncogene(s) located on chromosome 6.     

基于Haar特征和级联结构的CISH细胞检出

目的提出一种基于Haar特征和Gentle Adaboost级联结构的CISH细胞自动检出方法。方法根据乳腺癌CISH图像染色导致的细胞边缘模糊,不利于细胞的检出的问题。首先,对待检测图像进行光照补偿,减小光照对细胞检测造成的影响;然后,利用针对CISH细胞的Haar特征训练的级联强分类器快速检测细胞位置;最后,利用CISH细胞的结构特征对图像进行后处理,定位细胞的具体轮廓。结果实现乳腺癌CISH细胞的检出。结论实验结果表明,该算法检测速度快,并且具有较低误检率和漏检率,能够较好的应用于CISH细胞的快速检出。     

Association between CISH polymorphisms and spontaneous clearance of hepatitis B virus in hepatitis B extracellular antigen-positive patients during immune active phase

Background Some hepatitis B extracellular antigen （HBeAg）-positive chronic hepatitis B （CHB） patients in their immune active phase can clear the virus spontaneously and enter into an inactive hepatitis B virus （HBV） carrier state,indicating a benign prognosis.In this study,the association between cytokine-inducibie SRC homology 2 domain protein （C/SH） gene polymorphisms at-292 （rs414171） and the spontaneous clearance of HBV in HBeAg-positive CHB patients in immune the active phase was investigated.Methods Seventy HBeAg-positive CHB patients in the immune active phase were followed up for 76 weeks without antiviral therapy.The alanine transaminase,aspartate transaminase,HBV DNA,HBeAg and hepatitis B extracellular antibody levels were tested regularly.At week 76,27 patients were classified into group A （HBV DNA level below 2 104 IU/ml and the value of HBeAg declined below 10％ of the baseline at week 76）,and 43 patients were classified into group B （HBV DNA level higher than 2×104 IU/ml or the value of HBeAg did not decline substantially at week 76）.CISH （rs414171） polymorphisms were also tested using the iPLEX system.Results The HBV DNA levels at week 12 were significantly greater in group B compared with group A （group A：（6.87±1.40） log10IU/ml; group B：（7.61±1.38） log10IU/ml,P=0.034） and the HBeAg values were greater in group B at week 28 compared with group A （P=0.001）.The differences in HBV DNA and HBeAg values increased between the groups over time.Sixteen patients in group A and 11 in group B were genotype AA.Those with genotype AT or TT included 11 in group A and 31 in group B （AA vs.AT and TT,odds ratio 4.10 （95％ confidence interval：1.462-11.491）,P=0.006）.Conclusion CISH gene polymorphisms at-292 （rs414171） are associated with HBV clearance in HBeAg-positive CHB patients in the immune active phase,and AA is a favorable genotype for this effect.