A consensus multiplex PCR rCM-PCR)technique was devel-oped to detect high-risk fHPV 16/18), low-risk(HPV 6/11), and over 40 other types of human papillomavirus(HPV), separately but simultaneously. by mixing three pairs of consensus primers in the same PCR mixture, for ene amplification. Simultaneous detection of three groups 0f HPV DNA provides valuable infor-matfon for clinical practice and this procedure is simple and convenient for routine laboratory examinations. We detected HPV DNA sequences in plasmid HPV DNA and DNA extracted from tissues of condyloma acuminata and cervical carcinoma and from exfoliated cells of the lower genital tract of healthy Chinese women living in the People’s Republic of China. We confirmed that this simple, convenient, and cost-beneficial CM-PCR technique is reliable for the detection of HPV DNA se-quences.

To investigate the effects of human wild type p53 expression on the proliferation of cervical carcinoma cells, a plasmid, pM07 hp53, which contains a full-fength cDNA of the humao wild type p53 (wt p53) gene, was transfected into a cell line (TMCC 1) derived from an endocervical type, human papilloma virus positive adenocarcinoma of the uterine cervix. The exoge nous wt p53 expression induced growth suppression, morpho logical changes, and loss of anchorage independent growth of the tumor cells. As the wt p53 gene apparently plays a negative role in growth regulation of cervical carcinoma cells, this gene may possibly be of some use for treating subjects with a cervical carcinoma.

The standard telomeric repeat assay protocol (TRAP) was used to examine telomerase activity in 16 ovarian tumors, 16 cervical carcinomas, 4 uterine tumors, and 3 vaginal tumors. Telomerase activity was detected in 95% of these tumors, 88% of ovarian malignancies, and 100% of cervical, endometrial, and vaginal ma-lignancies. In contrast, telomerase activity was not evident in normal tissues or in benign proliferative lesions, such as leiomyomas,condyloma acuminata, and simple endometrial hyperplasia. These results suggest that telomerase activation is associated with im-mortalization or malignant transformation of gynecologic tumors.

目的 明确妊娠滋养细胞疾病和正常绒毛组织中的端粒酶表达状况，探索端粒酶活化在滋养细胞肿瘤形成中的作用。方法 应用以聚合酶链反应为基础的端粒酶测定方法（TRAP），对不同临床类型妊娠滋养细胞疾病及不同发育阶段正常妊娠绒毛组织共61例的端粒酶活性进行检测。结果 绒毛膜癌及侵蚀性葡萄胎36.4%（4/11）端粒酶阳性，葡萄胎26.3%（5/19）阳性；早孕60d以内绒毛全部（10/10）阳性，此期以后的胎盘组织（21/21）阴性。结论 端粒酶在妊娠早期以后表达受到抑制，其活化在滋养细胞肿瘤形成中可能起重要作用；其测定对此类疾病的诊断、预后和疗效评定有潜在价值。

Expression of the extracellular proteoglycan versican is elevated in a variety of human tumors, and this study aimed to investigate the potential mechanisms by which versican may affect tumor growth and angiogenesis.

Versican is a large chondroitin sulfate proteoglycan belonging to the lectican family. Alternative splicing of versican generates at least four isoforms named V0, V1, V2, and V3. We have shown that the versican V1 isoform not only enhanced cell proliferation, but also modulated cell cycle progression and protected the cells from apoptosis. Futhermore, the V1 isoform was able to not only activate proto-oncogene EGFR expression and modulate its downstream signaling pathway, but also induce p27 degradation and enhance CDK2 kinase activity. As well, the V1 isoform down-regulated the expression of the proapoptotic protein Bad. By contrast, the V2 isoform exhibited opposite biological activities by inhibiting cell proliferation and down-regulated the expression of EGFR and cyclin A. Furthermore, V2 did not contribute apoptotic resistance to the cells. In light of these results, we are reporting opposite functions for the two versican isoforms whose expression is differentially regulated. Our studies suggest that the roles of these two isoforms are associated with the subdomains CSβ and CSα, respectively. These results were confirmed by silencing the expression of versican V1 with small interfering RNA (siRNA), which abolished V1-enhanced cell proliferation and V1-induced reduction of apoptosis.

The papillomavirus E2 protein is required for viral transcriptional regulation, DNA replication and genome segregation. We have previously shown that the E2 transactivator protein and BPV1 genomes are associated with mitotic chromosomes; E2 links the genomes to cellular chromosomes to ensure efficient segregation to daughter nuclei. The transactivation domain of the E2 protein is necessary and sufficient for association of the E2 protein with mitotic chromosomes. To determine which residues of this 200-amino-acid domain are important for chromosomal interaction,E2 proteins with amino acid substitutions in each conserved residue of the transactivation domain were tested for their ability to associate with mitotic chromosomes. Chromatin binding was assessed by using immunofluorescence on both spread and directly fixed mitotic chromosomes. E2 proteins defective in the transactivation and replication functions were unable to associate with chromosomes, and those that were competent in these functions were attached to mitotic chromosomes. However, several mutated proteins that were defective for chromosomal interaction could associate with chromosomes after treatmentwith agents that promote protein folding or when cells were incubated at lower temperatures. These results indicate that precise folding of the E2 transactivation domain is crucial for its interaction with mitotic chromosomes and that this association can be modulated.