Establishment and Application of a Method for High-Risk Human Papillomavirus Genotyping in Cervical Cancer Tissue.

BACKGROUND: Persistent high-risk HPV infection is a major cause of cervical cancer and E6/E7 genes and the Li gene in the HPV genome are key targets to detect high-risk HPV. This study aims to explore the relationship between cervical lesions and E6/7 by establishing a polymerase chain reaction (PCR) to detect multiplex genes based on HPV EE7 genes. It is hoped that such methods will provide a more reliable method for clinical screening and the prevention of cervical cancer. METHODS: Based on alignment, specific primers were designed for HPV E6/E7 genes, the sequences of which came from five5 high-risk papillomaviruses that are common in China. This enabled an E6/E7 gene detection method based on multiplex PCR to be established. E6/E7 and Li gene testing were then performed on 65 cervical cancer tissue samples. The gene copy number of HPV E6/E7 genes and the Li gene were detected from different classifications by real-time fluorescence quantitative PCR. RESULTS: Out of the 65 cervical cancer tissue samples, 47 (72.31%) showed positive results in E6/E7 multiplex PCR, 21 (32.31%) showed positive results in the Ll gene PCR, and out of the 219 cervical exfoliate cell samples, 56 (25.57%) showed positive results in E6/E7 multiplex PCR, 21 (13.24%) showed positive results in the L1 gene PCR. There were significant differences (p < 0.05) between these two test results. Fluorescent quantitative PCR showed that the ratio of gene copy number of L1 genes and E6/E7 genes was below 1 (p < 0.05) in cervical cancer tissue, in which both the Li and E6/E7 genes coexist. CONCLUSIONS: The established HPV multiplex PCR assay based on the design of E6/E7 gene is a specific and sensitive method for the detection and genotype of five high-risk HPVs.

1-Phenyl-1-[(1-phenyl-ethyl)sulfonyl-methyl-sulfon-yl]ethane.

There are two mol-ecules in the asymmetric unit of the title compound, C(17)H(20)O(4)S(2). There are slight differences in the twist of the two rings relative to the S-C-S chain [dihedral angles of 48.41 (18) and 87.58 (16)° in the first mol-ecule and 45.98 (18) and 87.02 (18)° in the second] and the difference in the C-S-C-S torsion angles [176.68 (17) and -77.6 (2)° for the two independent mol-ecules].

Ethanol extract from Epimedium brevicornum attenuates left ventricular dysfunction and cardiac remodeling through down-regulating matrix metalloproteinase-2 and -9 activity and myocardial apoptosis in rats with congestive heart failure.

Epimedium, a traditional Chinese herb, has been used for the remedy of coronary heart disease, impotence and osteoporosis in traditional oriental medicine. However, despite extensive pharmacological studies, the molecular mechanism of the anti-heart failure effect of epimedium is little known. In the present study, we investigated the pharmacological action mechanism of ethanol extract of epimedium (EPI-ext) on isoproterenol-induced congestive heart failure (CHF) in rats. Isoproterenol administration resulted in severe heart failure, as shown by the increased levels of left ventricular (LV) weight index and heart rate, as well as LV end diastolic pressure, and by the decreased levels of LV systolic pressure, maximal rate of LV pressure rise, and maximal rate of LV pressure decline. EPI-ext dose-dependently reversed the changes of these cardiac morphometric and hemodynamic parameters. In addition, EPI-ext significantly inhibited the serum levels of tumor necrosis factor alpha, norepinephrine, angiotensin II and brain natriuretic peptide in rats with CHF and improved the histological changes including cadiocyte hypertrophy, cadiocyte degeneration, inflammatory infiltration, and cardiac desmoplasia. Furthermore, the expression and activities of matrix metalloproteinase-2 and -9, which regulate collagen production, were also blocked by EPI-ext. Moreover, myocardial apoptosis was remarkably attenuated by EPI-ext through the regulating Bcl-2/Bax axle. In conclusion, EPI-ext ameliorates LV dysfunction and cardiac remodeling through down-regulating matrix metalloproteinase-2 and -9 activity and myocardial apoptosis in rats with CHF.

[Significance of the high-intensity zone located in the posterior annulus fibrosus for diagnosing discogenic low back pain].

OBJECTIVE: To investigate the prevalence of high-intensity zone (HIZ) and the correlation between HIZ and low back pain (LBP). METHODS: The data of 1000 unselected cases, 566 males and 434 females, aged 49.49 (12 - 86), who underwent lumbar MRI were analyzed to examine the prevalence of HIZ. 200 of the 1000 cases with HIZ which had complete clinical history records, 112 males and 88 females, aged 52.29 (18 - 86), were divided into LBP (n = 115) and non-LBP series (n = 85). CT discography (CTD) was performed in 16 discs in 11 cases with HIZ and the correlation between the modified Dallas' discogram scale and pain reproduction in provocation discography was explored. RESULTS: HIZ located in posterior annulus fibrosus (AF) was shown in 378 discs in 317 cases, most located in L4/5 and L5/S1 (74.8%) and the location in left side (153 HIZs) was more common than that in right side (94 HIZs). HIZs were shown in superior region (15.4%), middle region (28.8%), as well as inferior region (55.8%) in sagittal plane. 115 of the 200 cases with HIZ who had complete clinical history records (57.5%) were symptomatic and 85 cases (42.5%) were asymptomatic. According to the modified Dallas' discogram scale, LBP could be provoked in 8 of the 9 patients with grade IV and could not be provoked in 6 of the 7 patients with the grade III. CONCLUSION: Whether HIZ is accompanied with LBP is related to the degree of disc degeneration. When CTD showed the degree of modified Dallas' grade IV and over most of the patients will show LBP. HIZ only indicates the possibility of discogenic LBP and can not replace provocation discography.

Chimerically fused antigen rich of overlapped epitopes from latent membrane protein 2 (LMP2) of Epstein-Barr virus as a potential vaccine and diagnostic agent.

Epstein-Barr virus (EBV) is prevalent throughout the world and is associated with several malignant diseases in humans. Latent membrane protein 2 (LMP2) of EBV plays a crucial role in the pathogenesis of EBV-associated tumors; therefore, LMP2 has been considered to be a potential immunodiagnostic and immunotherapeutic target. A multi-epitope-based antigen is a promising option for therapeutic vaccines and diagnoses of such malignancies. In this study, we systematically screened cytotoxic T lymphocyte (CTL), helper T cell (Th) and B-cell epitopes within EBV-LMP2 using bioinformatics. Based on the screen, two peptides rich in overlapping epitopes of both T cells and B cells were selected to construct a plasmid containing the sequence for a chimeric multi-epitope protein referred to as EBV-LMP2m, which is composed of LMP2aa195∼232 and LMP2aa419∼436. The EBV-LMP2m protein was expressed in E. coli BL21 (DE3) after prokaryotic codon optimization. Inoculation of the purified chimeric antigen in BALB/c mice induced not only high levels of specific IgG in the serum and secretory IgA in the vaginal mucus but also a specific CTL response. By using purified EBV-LMP2m as an antigen, the presence of specific IgG in the serum specimens of 202 nasopharyngeal carcinoma (NPC) patients was effectively detected with 52.84% sensitivity and 95.40% specificity, which represents an improvement over the traditional detection method based on VCA-IgA (60.53% sensitivity and 76.86% specificity). The above results indicate that EBV-LMP2m may be used not only as a potential target antigen for EBV-associated tumors but also a diagnostic agent for NPC patients.

[Application of MSCT and post-processing images to fractures of nasal bone in forensic identification].

OBJECTIVE: To evaluate the application of MSCT and post-processing images to fractures of nasal bone in forensic identification. METHODS: 134 cases were examined by thin slice scanning with MSCT and all of the data were sent to workstation for MPR and SSD. The result of MSCT was compared with that of X-ray. RESULTS: There are 55 (41.04%) cases of linear fracture, 46 (34.33%) cases of comminuted fracture, 27 (20.15%) cases of depressed fracture and 6 (4.48%) cases of no fracture in this sample. With X-ray or CR, 48 (35.82%) cases were misdiagnosed or underdiagnosed. 133 (99.25%) cases were confirmed by MSCT. Significance difference was found between X-ray and MSCT (chi2= 45.0816, P<0.001). CONCLUSION: MSCT and post-processing images might be the chief evidence for nasal fractures in forensic identification.

Multiple-integrations of HPV16 genome and altered transcription of viral oncogenes and cellular genes are associated with the development of cervical cancer.

The constitutive expression of the high-risk HPV E6 and E7 viral oncogenes is the major cause of cervical cancer. To comprehensively explore the composition of HPV16 early transcripts and their genomic annotation, cervical squamous epithelial tissues from 40 HPV16-infected patients were collected for analysis of papillomavirus oncogene transcripts (APOT). We observed different transcription patterns of HPV16 oncogenes in progression of cervical lesions to cervical cancer and identified one novel transcript. Multiple-integration events in the tissues of cervical carcinoma (CxCa) are significantly more often than those of low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL). Moreover, most cellular genes within or near these integration sites are cancer-associated genes. Taken together, this study suggests that the multiple-integrations of HPV genome during persistent viral infection, which thereby alters the expression patterns of viral oncogenes and integration-related cellular genes, play a crucial role in progression of cervical lesions to cervix cancer.

[Correlation of expression of nuclear factor kappaB, inhibitor protein kappaB, and Bcl-2 in cervical cancer to human papillomavirus infection].

BACKGROUND & OBJECTIVE: It was proved that nuclear factor kappaB (NF-kappaB) and its inhibitor protein kappaB (I-kappaB) played critical roles in regulating the expression of proapoptotic genes, and NF-kappaB is overexpressed in some tumors and related with tumorigenesis. However, the expression of NF-kappaB and I-kappaB in cervical cancer and its correlation to human papillomavirus (HPV) infection have not been reported yet. This study was to explore the correlation of the expression of NF-kappaB, I-kappaB, and Bcl-2 in cervical cancer to human papillomavirus (HPV) infection. METHODS: The expression of NF-kappaB, I-kappaB, and Bcl-2 were detected by immunohistochemistry in 46 specimens of cervical cancer and 26 specimens of normal cervical tissue. HPV DNA was detected by polymerase chain reaction (PCR). RESULTS: The positive rates of NF-kappaB and Bcl-2 were significantly higher in cervical cancer than in normal cervical tissue (60.9% vs. 23.1%, 52.2% vs. 0.0%, P < 0.01). The positive rate of I-kappaB was significantly lower in cervical cancer than in normal cervical tissue (30.4% vs. 57.7%, P < 0.05). The expression of NF-kappaB was closely related to Bcl-2 (P < 0.05). The positive rate of NF-kappaB was significantly higher in HPV DNA-positive cervical cancer than in HPV DNA-negative cervical cancer (81.3% vs. 35.7%, P < 0.05), but the positive rates of I-kappaB and Bcl-2 between these 2 groups had no significant difference (P >0.05). CONCLUSION: The high expression of NF-kappaB and Bcl-2 and the low expression of I-kappaB may be related to cervical carcinogenesis, and NF-kappaB expression may be related to HPV infection.