Expression and clinical significance of SSTR2, ADAM-12 and FLI-1 in small cell lung cancer tissue / 中国基层医药

Objective:To investigate the expression and clinical significance of somatostatin receptor 2 (SSTR2), a disintegrin and metalloproteinase-12 (ADAM-12), and friend leukemia virus integration-1 (FLI-1) in small cell lung cancer tissue.Methods:Eighty-two patients with small cell lung cancer who received treatment in Haiyang People's Hospital from January 2020 to January 2021 were included in this study. All patients underwent radical surgical resection. Small cell lung cancer tissues and adjacent tissues more than 2 cm from the edge of cancer tissues were harvested. The positive expression rates of SSTR2, ADAM-12, and FLI-1 in cancer tissues and adjacent tissues were determined by immunohistochemistry. The relationship between SSTR2, ADAM-12, FLI-1, and clinical characteristics were analyzed. The 1-year survival rate of patients with small cell lung cancer was calculated.Results:The positive rates of SSTR2, ADAM-12, and FLI-1 in small cell lung cancer tissue were 79.27% (65/82), 76.83% (63/82), and 78.05% (64/82), respectively, which were significantly higher than 19.51% (16/82), 17.07% (14/82), 20.73% (17/82) in the adjacent tissue ( χ2 = 58.57, 58.78, 53.90, all P < 0.05). SSTR2, ADAM-12, and FLI-1 were positively associated with lymph node metastasis, clinical stage, tissue invasion, tumor size, and histological grade (all P < 0.05). After controlling for gender, age, and others, SSTR2, ADAM-12, and FLI-1 were associated with lymph node metastasis, clinical stage, tissue invasion, tumor size, and histological grade (all P < 0.05). All patients were followed up for 1 year. Six patients were lost to follow-up. The 1-year survival rate of 76 patients with small cell lung cancer was 67.11% (51/76). The survival rate of patients with positive SSTR2, ADAM-12, and FLI-1 expression were lower than that of patients with negative SSTR2, ADAM-12, and FLI-1 expression ( χ2 = 3.93, 6.43, 7.52, all P < 0.05). Conclusion:SSTR2, ADAM-12, and FLI-1 are highly expressed in small cell lung cancer tissue. Combined detection of SSTR2, ADAM-12, and FLI-1 is conducive to the prognosis and evaluation of small cell lung cancer in patients. This study is innovative and scientific.

Effect of aluminum exposure on abnormal phosphorylationof tau protein in PC12 cells of rats / 预防医学

Objective @#To investigate the effect of aluminum exposure on expression of miR-497-5p, wingless murine breast cancer virus integration site family member 3a (Wnt3a), β-catenin protein, glycogen synthase kinase-3β (GSK-3β) protein and tau protein in rat adrenal pheochromocytoma PC12 cells, so as to provide insight into unraveling the mechanisms underlying aluminum exposure-induced abnormal phosphorylation of tau protein.@* Methods@# PC12 cells were exposed to Al(mal)3 at concentrations of 0, 100, 200, 400 μmol/L for 24 h. The viability of PC12 cells was measured using cell counting kit-8 (CCK-8) assay. The relative expression of miR-497-5p and Wnt3a was detected using a real-time fluorescent quantitative PCR (RT-qPCR) assay, and the expression of Wnt3a, β-catenin, GSK-3β, P-GSK-3β (Ser9), tau and p-tau (Ser396) proteins were determined using Western blotting. @*Results @#The viability of PC12 cells appeared a tendency towards a decline with the increase of aluminum dose (Ftrend=323.473, P=0.001). RT-qPCR assay detected that the relative miR-497-5p expression appeared a tendency towards a rise with the increase of aluminum dose (Ftrend=14.888, P=0.031), and the relative Wnt3a expression appeared a tendency towards a decline with the increase of aluminum dose (Ftrend=165.934, P<0.001). The miR-497-5p expression negatively correlated with the relative Wnt3a expression (r=-0.693, P=0.012). The expression of Wnt3a (Ftrend=357.656, P=0.001), β-catenin (Ftrend=208.750, P=0.001) and p-GSK-3β (Ser9) proteins (Ftrend=512.583, P<0.001) appeared a tendency towards a decline with the increase of aluminum dose, and the expression of GSK-3β (Ftrend=39.965, P<0.001), tau (Ftrend=277.929, P=0.006) and p-tau (Ser396) proteins (Ftrend=96.247, P=0.002) appeared a tendency towards a rise with the increase of aluminum dose. @*Conclusion@# Up-regulation of miR-497-5p and GSK-3β expression and down-regulation of Wnt3a and β-catenin expression may be a mechanism underlying aluminum exposure-induced abnormal phosphorylation of tau protein.

In silico analysis of genomic variables associated to HPV16 integration sites / Análisis in silico de variables genómicas asociadas a sitios de integración de HPV16

Background: Despite current prophylactic interventions, a significant proportion of patients suffers a cancer-specific mortality, leading to a global awareness of the importance of identifying factors associated to the etiology of HPV-associated cancer. According to this, HPV-DNA integration into human genome is an important event in the pathogenesis. Purpose: To identify in silico, molecular regions of the genome where the HPV integration events occur Methods: We performed a bioinformatic study based on a systematic search in Medline through PubMed, Embase and Lilacs from inception to April 2019. We used the UCSC Genome Browser Home (https://genome.ucsc.edu) to evaluate the genetic environment. Results: HPV integration sites by anatomical location related to cervical cancer were 374 (61%). In addition, 325 (87%) of these integration sites had HPV-16, 21 (5%) had HPV-18 and 28 (7%) had another type of genotype. Oro-pharyngeal cavity was the second anatomic site with 162 (26%) integration sites. It is noteworthy that the HPV-16 was found integrated into 160 (99%) analyzed sites. Conclusion: Our results suggest that many of the integration sites reported in the scientific literature are HPV 16 from squamous cell carcinomas and 50% of HPV16 were integrated into transcriptional units that might affect the expression of gene target.

Antecedentes: A pesar de las intervenciones profilácticas actuales, una proporción significativa de pacientes muere debido al cáncer, lo que aumenta la conciencia global de la importancia de identificar los factores asociados a la etiología del cáncer asociado al VPH. Según esto, la integración del ADN-VPH en el genoma humano es un evento importante en la patogénesis. Propósito: Identificar in silico, las regiones moleculares del genoma donde ocurren los eventos de integración del VPH Métodos: Realizamos un estudio bioinformático basado en una búsqueda sistemática en Medline a través de PubMed, Embase y Lilacs desde el inicio hasta abril de 2019. Utilizamos el UCSC Genome Browser Home (https://genome.ucsc.edu) para evaluar el entorno genético. Resultados: Los sitios de integración del VPH relacionados con el cáncer de cuello uterino fueron 374 (61%). Además, 325 (87%) de estos sitios de integración tenían VPH-16, 21 (5%) tenían VPH-18 y 28 (7%) tenían otro tipo de genotipo. La cavidad orofaríngea fue el segundo sitio anatómico con 162 (26%) sitios de integración. Es de destacar que el VPH-16 se encontró integrado en 160 (99%) sitios analizados. Conclusión: Nuestros resultados sugieren que muchos de los sitios de integración reportados en la literatura científica que presentan al VPH-16 son carcinomas de células escamosas y que el 50% de estos VPH-16 se integraron en unidades transcripcionales que podrían afectar la expresión de algún gen objetivo.

Integration, Viral Charge and E2 mRNA Levels in the Progresion of Intraepitelial Cervical Lesions / Integración, carga viral y niveles de arn mensajero de e2 de vph 16 en la progresión de lesiones intraepiteliales cervicales

RESUMEN Entre las lesiones intraepiteliales escamosas cervicales (LIE) es importante distinguir aquellas asociadas con mayor riesgo de cáncer de cuello uterino. El objetivo de este trabajo fue evaluar si los niveles de expresión de E2 del VPH16 en mujeres con LIE y con evidencia de integración viral se asocian con el grado de la lesión. Se analizaron 109 cepillados cervicales positivos para VPH 16 provenientes de 19 mujeres sin LIE, 45 mujeres con LIE de bajo grado (LIEBG) y 45 mujeres con LIE de alto grado (LIEAG). Se cuantificó el número de copias de ARNm de E2 y de los genes E2 y E6 mediante PCR en tiempo real para determinar la carga viral (E6) y la proporción E2/E6 para evaluar la integración viral. Se encontraron frecuencias similares de expresión de E2 en LEIBG y LEIAG 15/45 (33 %), la frecuencia en mujeres sin lesión fue menor 3/19 (15,8 %), todos los casos en los que se observó expresión del gen E2 tenían mezcla de ADN viral episomal e integrado. La carga viral aumentó significativamente a mayor grado de la lesión (ρ =0,049), mientras que la proporción E2/E6 disminuyó (ρ=0,049). El análisis ROC mostró una baja capacidad de los tres parámetros virales para distinguir entre lesiones de bajo y alto grado. En conclusión, aunque las lesiones con presencia de ADN viral mixto e integrado y expresión de E2 podrían estar en menor riesgo de progresión, y la carga viral y la integración se relacionaron con mayor gravedad de la lesión, su valor clínico como biomarcadores de LEIAG es limitado.

ABSTRACT It is important to distinguish among squamous intraepithelial lesions (SIL) those associated with increased risk cervical cancer. Our aim was to evaluate if the expression level of gen E2 in women with SIL and evidence of viral integration is associated to the grade of lesion. Cervical scrapes HPV16 positive from 19 women with normal histology, 45 women with low-grade SIL (LSIL) and 45 women with high-grade SIL (HSIL) were analyzed. Real-time PCR was used to quantify the mRNA of E2 and E2 and E6 genes to calculate viral load (E6) and the ratio E2/E6 to assess viral integration. Similar frequencies of E2 expression were found in LSIL and HSIL15/45 (33 %), the frequency in women without SIL was lower 3/19 (15.8 %), and all cases with E2 gene expression had mixed episomal and integrated viral DNA. The viral load increased significantly with the grade of SIL (ρ= 0.049), while E2/E6 ratio decreased (ρ=0.049). The ROC analysis showed low capacity of the three viral parameters analyzed to distinguish between low and high grade SIL. In conclusion although SIL with mixed and integrated viral DNA with E2 expression could be at lower risk of progression, and viral load and integration were associated with higher severity of the lesion, its clinical value as biomarkers of HSIL is limited.

Genotipificación, niveles de expresión y estado físico del virus del papiloma humano en pacientes colombianos con cáncer de células escamosas en la cavidad oral / Genotyping, levels of expression and physical status of human papilloma virus in oropharyngeal squamous cell carcinoma among Colombian patients

Introducción. Uno de los factores de riesgo del carcinoma de células escamosas en la cavidad oral es la infección por el virus del papiloma humano (HPV), cuyas prevalencias dependen de la región geográfica. Objetivo. Identificar los tipos del virus del papiloma humano más frecuentes en el cáncer de la cavidad bucal, sus niveles de expresión y el estado físico del genoma viral. Materiales y métodos. Se seleccionaron 46 pacientes que asistían a los servicios de cirugía de cabeza y cuello en Bogotá, Manizales y Bucaramanga. El examen histopatológico de las muestras incluidas en el estudio demostró la presencia de carcinoma de células escamosas en la cavidad oral en todas ellas. Se extrajo el ADN para genotipificar el virus y determinar el estado físico de su genoma, y el ARN para determinar los transcritos virales mediante reacción en cadena de la polimerasa en tiempo real. Resultados. La prevalencia del virus del papiloma humano en los tumores fue de 21,74% (n=10) y el tipo viral más frecuente fue el HPV-16 (nueve casos). La expresión viral del HPV-16 fue baja (una de 11 copias) y el estado físico predominante fue el mixto (ocho casos), con prevalencia de la disrupción en el sitio de unión de E1 y E2 (2525 a 3720 nucleótidos). Conclusión. En los pacientes con carcinoma de cavidad oral incluidos en este trabajo, la frecuencia del virus del papiloma humano fue relativamente baja (21,7 %) y el tipo viral más frecuente fue el HPV-16, el cual se encontró en forma mixta y con baja expresión de E7 , lo cual puede ser indicativo de un mal pronóstico para el paciente.

Introduction: One of the risk factors for squamous cell oropharyngeal carcinoma is infection with the human papilloma virus (HPV), with prevalences that vary depending on the geographical region. Objective: To identify the most frequent HPV viral types in oropharyngeal cancer, the levels of expression and the physical condition of the viral genome. Materials and methods: Forty-six patients were included in the study from among those attending head and neck surgical services in the cities of Bogotá, Manizales and Bucaramanga. In the histopathological report all study samples were characterized as oropharyngeal squamous cell carcinoma. DNA extraction was subsequently performed for HPV genotyping and to determine the physical state of the viral genome, as well as RNA to determine viral transcripts using real-time PCR. Results: HPV prevalence in tumors was 21.74% (n=10) and the most common viral type was HPV-16 (nine cases). Viral expression for HPV-16 was low (one of 11 copies) and the predominant physical state of the virus was mixed (eight cases), with disruption observed at the E1 - E2 binding site (2525 - 3720 nucleotides). Conclusion: The prevalence of HPV associated with oropharyngeal carcinoma among the Colombian study population was 21.7%, which is relatively low. The most frequent viral type was HPV-16, found in a mixed form and with low expression of E7 , possibly indicating a poor prognosis for these patients.

Changes and significance of B cell specific monoclonal murine leukemia virus integration site -1 and helper T cell-related cytokine in children with immune thrombocytopenia / 中华实用儿科临床杂志

Objective To analyze the functional changes and the clinical significance of B cell specific mono-clonal murine leukemia virus integration site -1 (Bmi -1 )and Th1 /Th2 cells in children with newly diagnosed im-mune thrombocytopenia(ITP)by testing the mRNA expressions of Bmi -1,helper T cell -related cytokine interferon (IFN)-γand interleukin(IL)-4 in children with newly diagnosed ITP.Methods Thirty -six cases of patients with newly diagnosed ITP in the experimental group came from the inpatient and outpatient children admitted to the Depart-ment of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from April to December 201 3.In the control group,26 cases of children requiring selective operation were admitted to the Department of Pediatric Surgery during the same period.The mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes were detected by means of the reverse transcription -polymerase chain reaction(RT -PCR)method,and were analyzed and compared by t test and linear correlation analysis.Results (1 )The mRNA expressions of Bmi -1,IFN -γand IL -4 in peripheral blood lymphocytes in the experimental group were 2.63 ±0.54,3.84 ±0.43 and 1 .44 ±0.39,respec-tively;while the mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes in the control group were 3.91 ±0.92,2.88 ±0.57 and 1 .87 ±0.34,respectively.The levels of IFN -γof the experimental group were significantly higher than those of the control group (P 0.05).Conclusions Bmi -1 may be involved in the pathogenesis of ITP by regulating Th cell, and Th cell dysfunction may occur in the children with ITP,and the disproportion between Th1 and Th2 may be due to the advantages of Th1 .

Murine leukemia virus-derived retroviral vector has differential integration patterns in human cell lines used to produce recombinant factor VIII

OBJECTIVE: Nowadays recombinant factor VIII is produced in murine cells including in Chinese hamster ovary (CHO) and baby hamster kidney cells (BHK). Previous studies, using the murine leukemia virus-derived retroviral vector pMFG-FVIII-P140K, modified two recombinant human cell lines, HepG2 and Hek293 to produce recombinant factor VIII. In order to characterize these cells, the present study aimed to analyze the integration pattern of retroviral vector pMFG-FVIII-P140K. METHODS: This study used ligation-mediated polymerase chain reaction to locate the site of viral vector integration by sequencing polymerase chain reaction products. The sequences were compared to genomic databases to characterize respective clones. RESULTS: The retroviral vector presented different and non-random profiles of integration between cells lines. A preference of integration for chromosomes 19, 17 and 11 was observed for HepG2FVIIIdB/P140K and chromosome 9 for Hek293FVIIIdB/P140K. In genomic regions such as CpG islands and transcription factor binding sites, there was no difference in the integration profiles for both cell lines. Integration in intronic regions of encoding protein genes (RefSeq genes) was also observed in both cell lines. Twenty percent of integrations occurred at fragile sites in the genome of the HepG2 cell line and 17% in Hek293. CONCLUSION: The results suggest that the cell type can affect the profile of chromosomal integration of the retroviral vector used; these differences may interfere in the level of expression of recombinant proteins. .

Role of hepatitis b virus integration in evolution of hepatocellular carcinoma / 第二军医大学学报

Hepatitis B virus (HBV)-human DNA integration is a commonly seen event in the evolutionary process of hepatocellular carcinoma (HCC). HBV chronic infection induces HBV mutation, HBV integration and the inflammatory microenvironment alternation in the hosts, which provides a evolutionary soil for the hepatocarcinogenesis. The second generation sequencing technique provides a better approach for understanding the HBV integration mechanism in HCC. This review focuses on the features of HBV integration and influences of HBV integration on human, virus genomes, and discusses the association of HBV integration with HCC.

Preliminary study on change of immunologic factors within cervical microenvironment infected with HPV 16, 58 and integrated state of virus in cervical cells / 中国免疫学杂志

Objective:To investigate the relationship between changes of the concentration of IL -12,IL-4,IFN-γwithin cervical local immune microenvironment and cervix infected with HPV 16,58,to understand the prevalence of physical status of HPV 16/58 DNA into the host genome in cervical lesions of different grades and to detect the relation between the integrated state of virus and the progress of cervical lesions.Methods: Detecting 89 cases of cervicovaginal lavage fluid and 140 cytological specimens of the cervix which from patients having had HPV-DNA genotyping assays and coming from the Affiliated Tumor Hospital of Guangxi Medical University from 2012.8 to 2013.5.89 cases of cervicovaginal lavage fluid were divided into 58 cases of HPV16 (+) and 31 cases of HPV58(+).Cytokines IL-12, IL-4, IFN-γin the cervical microenvironment were detected by ELISA assay.140 cases of cervical exfoliated cells were divided into 106 cases of HPV16(+) and 34 cases of HPV58(+).Using qRT-PCR to quantify the copy number of E2 and E6 genes and analyze the physical status of HPV 16/58 DNA according to E2/E6.Results:①The level of IL-4 in HPV16(+) was significantly higher than that in HPV58(+)(P<0.05).The levels of IL-12 and IFN-γwere lower than that in HPV58(+)(P<0.05).②Significant differences of DNA state were found between the groups of HPV 16(+) and HPV58(+)(P<0.05)and the rate of HPV16 DNA integration was significantly larger than that of HPV 58 DNA.Conclusion:The infection of HPV16 and HPV58 not only can lead to reduction of IL-12,IFN-γand increasing of IL-4 in cervical local immune microenvironment but also lead to viral integration in cervical cells,resulting in cervical lesions and cervical cancer through the internal and external changes in cervical cells .HPV16 is more likely to cause cervical cancer tissue than 58.

Epidemiología genómica y paraparesia espástica tropical asociada a la infección por el virus linfotrópico humano de células T tipo 1 / Genome epidemiology and tropical spastic paraparesis associated with human T-cell lymphotropic virus type 1

OBJETIVO: Caracterizar el ambiente genómico de las secuencias adyacentes al virus linfotrópico humano de células T tipo 1 (HTLV-1) en pacientes con paraparesia espßstica tropical y mielopatía asociada a la infección con HTLV-1 (PET/MAH) de diferentes regiones de Colombia y del Japón. MÉTODOS: Se enfrentaron 71 clones recombinantes con secuencias del genoma humano adyacentes al 5'-LTR de pacientes con PET/MAH, a las bases de datos del Genome Browser y del Gen-Bank. Se identificaron y analizaron estadísticamente 16 variables genómicas estructurales y composicionales mediante el programa informßtico R, versión 2.8.1, en una ventana de 0,5 Mb. RESULTADOS: El 43,0 por ciento de los provirus se localizaron en los cromosomas del grupo C; 74 por ciento de las secuencias se ubicaron en regiones teloméricas y subteloméricas (P < 0,05). Un anßlisis de conglomerados permitió establecer las relaciones jerßrquicas entre las características genómicas incluidas en el estudio; el anßlisis de componentes principales identificó las componentes que definieron los ambientes genómicos preferidos para la integración proviral en casos de PET/MAH. CONCLUSIONES: El HTLV-1 se integró con mayor frecuencia en regiones de la cromatina ricas en islas de citocina fosfato guanina (CpG), de alta densidad de genes y de repeticiones tipo LINE (elemento disperso largo [long interspersed element]) y transposones de ADN que, en conjunto, conformarían los ambientes genómicos blanco de integración. Este nuevo escenario promoverß cambios sustanciales en el campo de la salud pública y en el manejo epidemiológico de las enfermedades infecciosas, y permitirß desarrollar potentes herramientas para incrementar la eficiencia de la vigilancia epidemiológica.

OBJECTIVE: Characterize the genomic environment of the sequences adjacent to human T-cell lymphotropic virus type 1 (HTLV-1) in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) in different regions of Colombia and Japan. METHODS: A total of 71 recombinant clones with human genome sequences adjacent to 5' LTR in patients with HAM/TSP were compared to the Genome Browser and GenBank databases. Sixteen structural and compositional genome variables were identified, and statistical analysis was conducted in the R computer program, version 2.8.1, in a 0.5 Mb window. RESULTS: A total of 43.0 percent of the proviruses were located in the group C chromosomes; 74 percent of the sequences were located in the telomeric and subtelomeric regions (P < 0.05). A cluster analysis was used to establish the hierarchical relations between the genome characteristics included in the study. The analysis of principal components identified the components that defined the preferred genome environments for proviral integration in cases of HAM/TSP. CONCLUSIONS: HTLV-1 was integrated more often in chromatin regions rich in CpG islands with a high density of genes and LINE type repetitions, and DNA transposons which, overall, would form the genomic environments targeted for integration. This new scenario will promote substantial changes in the field of public health and in epidemiological management of infectious diseases. It will also foster the development of powerful tools for increasing the efficiency of epidemiological surveillance.

HPV type 16 cervical infection in eastern Guangzhou and viral gene analysis / 中华微生物学和免疫学杂志

Objective To investigate the distribution of human papillomavirus (HPV) type 16 in women cervical infection in eastern Guangzhou, polymorphism of E6/E7 gene and association of gene dosage with disease progression. Methods Flow-through hybridization and gene chips were applied in HPV sub-type identification to screen out HPV-16 positive samples from cervical epithelium samples. HPV-16 E6/E7 gene was amplified through PCR with specific primers. The PCR products were cloned into pMD18-T vectors and fragments were determined through sequencing. Polymorphism analysis were performed through align-ment tools. Fluorescence quantitive PCR were used for the detection of viral E6 gene and L1 gene. Results Thirty-six (4.5%) HPV-16 positive samples were screened out through flow-through hybridization from 806 cervical epithelium samples. HSIL and above happened in 18 (50.0%) of the 36 HPV-16 positive patients. Within E6/E7 gene sequences from 7 selected samples, we found 15 sites with variances and 8 of them would cause coding amino acid change. HIL group (A, 11 cases) and LSIL group (B, 14 cases) possess significantly different gene dosage of both viral E6 gene and LI gene (P <0.05). The ratios of L1/E6 be-tween the 2 groups was not significantly different(P=0.19). Conclusion HPV-16 cervical infection oc-curs in 4.5% women (17-62 years old) in eastern Guangzhou. HIL or above accompany with half of the HPV 16 infected women. Viral load is probably associated with cervical HSIL, though L1/E6 ratios do not suggest viral integration.

Detection of hepatitis B virus DNA integration in hepatocellular carcinoma using hepatitis B virus-Alu-polymerase chain reaction / 中华传染病杂志

Objective To detect the integration of hepatitis B virus (HBV) DNA in HBVrelated human hepatocellular carcinomas (HCC). Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction (PCR) with specially designed U-base primers. According to the known genes and human Alu repeat sequences (Alu repeat) , primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies.Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information (NCBI) basic local alignment search tool (BLAST) and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn't show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

Integration of HPV 16/18 DNA in Vulvar Intraepithelial Neoplasia / 中华皮肤科杂志

Objective To investigate the pathogenic role of high risk HPVs and genomic integration of HPV 16/18 DNA in vulvar intraepithelial neoplasia (VIN) patients. Methods Hybrid Capture HPV DNA assay and PCR were performed to detect high-risk HPVs and HPV 16/18 positive samples. Then RT-PCR, nested PCR and Southern blotting analysis were used in HPV 16/18 positive samples for the amplification and analysis of papillomavirus oncogene transcripts. Results In 32 cases of VIN patients, 24(75%) high-risk HPVs were detected, 23 were HPV 16, and only 1 was HPV 18. Genomic integration of HPV 16/18 was observed in 8 cases of VIN Ⅲ (7 HPV 16 and 1 HPV 18). Except 1 case of VIN Ⅱ, 15 of 23 HPV-16 positive specimens displayed HPV16 episomal transcripts, 7 displayed integrated transcripts. Conclusions HPV 16 is positive in most cases of VIN Ⅱ and VIN Ⅲ. Integration of HPV 16/18 oncogene only occurs in VIN Ⅲ. It is concluded that the integration of high-risk HPVs oncogene is related with the occurrence, and the development to vulvar cancer, of VIN.