Multiple Skin Cancers in a Renal Transplant Recipient: A Patient Report with Analyses of Human Papillomavirus and Human Polyomavirus Infection

Skin cancer is an important complication in renal transplant recipients. Associations of transplant-related skin tumor with ultraviolet radiation, age at transplantation, type of immunosuppressant drug administered, and viral infection have been reported; however, the details remain unclear. We report a 61-year-old man who had underwent renal transplantation at 38 years of age and developed multiple skin tumors or squamous cell carcinomas (SCCs). Polymerase chain reaction (PCR) analyses of the patient’s 12 tumors for viral DNAs of cutaneous or mucosal human papillomavirus (HPV) and 6 human polyomaviruses (MCPyV, trichodysplasia spinulosa-associated, BK, JC, KI and WU polyomaviruses) only detected cutaneous HPV-DNA in only 5 of the tumors; no other viruses were detected. Real-time PCR showed high loads of cutaneous HPV in 3 SCCs and very low loads of MCPyV in 9. Immunohistochemistry revealed no tumor cell expression for MCPyV-large T-antigen or mucosal HPV. Our report not only reconfirmed the association of cutaneous HPV5 with skin cancer in renal transplant recipients in previous studies but also showed no relevant association of 6 human polyomaviruses and mucosal HPV with skin tumors.     

Human Papillomavirus Infection Rates before and after the Introduction of Prophylactic Vaccines in Kunming,Yunnan, China

Purpose: The purpose of this study was to determine the prevalence of human papillomavirus (HPV) in the population in Kunming, Yunnan, China, before and after the introduction of HPV preventive vaccines. Materials and Methods: In total, 28,959 patients were enrolled in this study between 1 January 2016 and 31 August 2019. HPVs were genotyped using a flow-through hybridisation technique, and differences in HPV infection rates before and after the introduction of an HPV vaccine were determined. Results: The prevalence of HPV before and after the introduction of HPV vaccines was 17.74% and 17.11%, respectively. This difference was not statistically significant (χ² = 1.920, P > 0.05). The HPV infection rates showed a bimodal U-shaped curve for all age groups. The most common genotypes of HPV detected were HPV52, HPV16, HPV58 and HPV39. Conclusion: Although the overall HPV infection rate in Kunming did not change significantly after the introduction of HPV vaccines, differences in HPV infection rates and multi-typic HPV infection rates were evident in certain age groups.     

湖北省人乳头瘤病毒基因分型检测分析

目的 调查湖北省女性人群人乳头状瘤病毒(human papillomavirus,HPV)感染状况,为宫颈癌防治提供参考依据.方法 收集2013年1月至2014年11月在湖北省中山医院就诊的3956例宫颈脱落细胞标本,采用PCR体外扩增和DNA反向点杂交相结合的DNA芯片技术,对宫颈脱落细胞标本进行HPV基因分型检测.结果 3956例患者中共检出HPV阳性818例,阳性率为20.68％,其中高危型HPV感染阳性率为15.77％(624/3956).在被检测的23个HPV-DNA亚型中,最常见的依次为16型(3.72％,147/3956),52型(3.67％,145/3956),43型(3.64％,144/3956),58型(2.78％,110/3956),18型(1.49％,59/3956),33型(1.06％,42/3956),未检出MM4型.HPV阳性者中多重感染率为17.36％(142/818),以二重感染最常见(85.21％,121/142).结论 湖北省地区高危型HPV感染以HPV16、52、58、18和33比较常见,低危型HPV感染以HPV43比较常见.     

Human Papillomavirus Prevalence and Genotype Distribution among Turkish Women with or Without Cervical Lesion

Context: Human papillomavirus (HPV) infection is the main cause of cervical cancer, but the risk is associated with the various HPV genotypes which may be found in women with or without clinical findings. Aims: We aimed to identify HPV prevalence and genotype distribution in women with or without cervical lesions admitted to Gynaecology and Obstetrics Clinics of one of the largest private hospitals in Istanbul between 2013 and 2017. Subjects and Methods: In the present study, cervical cytobrush samples collected from 2464 women with different cytological conditions, and investigated for the presence of HPV, and the different genotypes. Results were evaluated based on the HPV positivity in different cytological findings, and ages. Furthermore, distribution of high-risk (HR) and low-risk (LR) genotypes in different groups was investigated. Results: Among all participants, 1925 (78.1%) was with the normal cytological condition, 354 (14.4%) with ASC-US; 151 (6.1%) with low-grade squamous intraepithelial lesion (LSIL), and 34 (1.4%) with high-grade squamous intraepithelial lesion (HSIL). Our results showed that 649 out of 2464 patients (26.3%) were positive, and 1815 (73.7%) were negative for the presence of HPV. Among 649 positive patients, 223 (34.3%) were found positive for more than one genotype. HPV 16 was found the most common HR-HPV type in ASC-US and LSIL whereas HPV 18 was the most common in HSIL. HPV 6 was found the most common LR-HPV type in ASC-US and LSIL whereas HPV 11 was the most common in HSIL. 26.9% of women <50 years old, and 22.3% of women ≥50 years old was positive for HPV. The most common HR-HPV genotype was 16 in both groups with (19%) or without (17%) abnormal cytology. Conclusions: We concluded that HPV prevalence and genotype distribution in women with or without clinical findings is an important predictor of cervical cancer.     

Discovery and characterization of potent small molecule inhibitors of the high affinity proline transporter

The mammalian proline transporter (PROT) is a high affinity Na⁺/Cl⁻-dependent transporter expressed in specific regions of the brain. It is homologous to other neurotransmitter transporters such as glycine, norepinephrine, serotonin, and dopamine transporters. PROT is enriched in glutamatergic synaptic terminals and may play an important role in the regulation of excitatory neurotransmission. No non-peptide small molecule inhibitors have been described for this transporter. To study its physiological role in the central nervous system and evaluate its potential as a therapeutic target, we established cell lines that stably express recombinant hPROT and characterized its kinetic properties for proline uptake. We then screened for inhibitors and identified a series of compounds that inhibit hPROT-mediated proline uptake. A known compound, benztropine, was found to inhibit hPROT with an IC₅₀ of 0.75 μM. A series of novel compounds were also found, one of which, LP-403812, showed an IC₅₀ of approximately 0.1 μM on both recombinant human and mouse PROT without significant inhibition of glycine and dopamine transporters at concentrations up to 10 μM. This compound also inhibited proline transporter activity of mouse brain synaptosomes with the same potency. These inhibitors provide important tools for the understanding of PROT functions in the brain and may lead to the development of therapeutic agents for certain neurological disorders.     

北京市北苑地区女性HPV感染现状及基因型分布研究

目的 调查我院辖区范围内即北京市北苑地区成年女性人乳头状瘤病毒(HPV)感染现状及亚型分布情况.方法 采用核酸分子快速导流杂交基因芯片分型技术对本院辖区范围内(北苑地区)来我院妇科门诊就诊的14582例女性患者采集宫颈脱落细胞标本进行HPV及21种亚型检测.结果 ①HPV感染阳性检出2419例,感染阳性率16.59％(2419/14582),在HPV感染阳性人群中,以单一高危型HPV感染为主,单一感染HPV阳性1951例,占HPV阳性感染人群80.65％(1951/2419);多重感染以二重感染为主,多重感染HPV阳性468例,二重感染HPV阳性361例,占HPV阳性感染人群14.92％(361/2419),以高危型感染以为主.②各HPV基因亚型检出总和为3035项次,高危型HPV感染占91.73％(2784/3035),排在前五位由高到低依次是HPV16(17.36％,527/3035)、HPV52(11.89％,361/3035)、HPV58(11.80％,358/3035)、HPV53(8.27％,251/3035)、CP8304(7.05％,214/3035),以HPV16型为主;低危型HPV感染占8.27％(251/3035),由高到低依次是HPVl1 (3.95％,120/3035)、HPV6(3.57％,108/3035)、HPV44(0.43％,13/3035)、HPV42 (0.16％,5/3035)、HPV43 (0.16％,5/3035),以HPV11为主.③按年龄段划分,≤20岁年龄组阳性率最高,阳性率31.30％ (36/115);其次是≥61岁年龄组,阳性率21.05％ (64/304).结论 本院辖区范围内(北苑地区)成年女性HPV感染阳性率16.59 ％(2419/14582),以单一高危型HPV感染为主,高危型HPV16、HPV52、HPV58、HPV53、CP8304是本院辖区范围内成年女性HPV感染的主要亚型.     

人乳头瘤病毒基因型在湘潭地区女性下生殖道感染中的分布特点

了解人乳头瘤病毒(HPV)基因型在本地区女性下生殖道感染的分布及特点.采用快速分子导流杂交分型技术对2009年5月至2010年6月间本院妇产科就诊的2682例女性进行生殖道21种HPV分型检测.结果显示:HPV阳性率为23.8%(639/2682),21种HPV亚型均有检出,其中单一型感染率为18.3%(490/268...     

Depressive Symptoms and Cervical Neoplasia in HIV+ Low-Income Minority Women with Human Papillomavirus Infection

Background  Prior work has related elevated life stress to greater risk of cervical neoplasia in women with human immunodeficiency virus (HIV) and human papillomavirus (HPV). Purpose  This study investigated associations between depressive symptoms and cervical neoplasia in HIV+ HPV+ women. Participants were 58 HIV+ HPV+ women. Method  Participants underwent colposcopy, including HPV screening, Papanicolaou smear, and cervical biopsy to determine study eligibility. Eligible participants completed the Beck Depression Inventory (BDI) and the Center for Epidemiologic Studies-Depression (CES-D) scale. Results  Presence and severity of clinically significant depressive symptomatology were associated with cervical neoplasia. Hierarchical logistic regression analysis revealed that women with greater depressive symptoms had marginally greater odds of presenting with cervical neoplasia (BDI: OR = 1.16, p = 0.092; CES-D: OR = 1.15, p = 0.067. Women with greater somatic depressive symptoms, specifically, had significantly greater odds of presenting with cervical neoplasia (BDI: OR = 1.86, p = 0.027; CES-D: OR = 1.56, p = 0.017). Conclusion  These findings suggest that screening HIV+ women for somatic depression may help identify those at risk for cervical neoplasia. Future depression research with medical populations should discern somatic depressive symptoms from disease symptoms, as they may have important value in independently predicting health outcomes.     

子宫颈癌中人乳头瘤病毒16型E7基因检测及变异分析

用聚合酶链反庆法检测宫颈癌中人乳头瘤病毒１６型转化基因Ｅ７，检出阳性率为１０／２２，其中４例进行ＰＣＲ－ＳＳＣＰ分析发现该基因与标准对照株存在明显变异。核苷酸序列分析证实有两处发生碱基突变。研究提示，ＰＣＲ方法能快速检测宫颈癌中人乳头瘤 转化基因，可作为宫颈癌的辅助诊断方法，ＳＳＣＰ分析能发现病毒株间小变异的基因片段。     

人乳头瘤病毒检查与影响因素的探讨

使用DNA基因杂交捕获二代法(hc2)检测3273例宫颈外口及宫颈管分泌物人乳头瘤病毒(HPV),并探讨试验中影响因素。结果表明:HPV阳性率为28.0%,而年龄在18～45岁阳性率为31.6%。随着年龄增加HPV阳性率则呈下降趋势。hc2是宫颈癌筛查最佳方案中的方法之一。45批实验中,42批实验低危型和高危型HPV质控、阴性质控和校准品的各种参数均在试剂盒给出的范围内,结果数据可靠。在三批失败的实验中,因质控的参照品未与待测样品在相同条件下水解,操作者加样的准确性、重复性差和样本被污染而失败。建立稳定的hc2方法为临床提供可靠的数据至关重要。     

P16 overexpression and human papillomavirus infection in small cell carcinoma of the uterine cervix

Carcinogenesis of cervical cancer has been investigated, and p16(INK4a) overexpression in squamous cell carcinoma of the cervix has been reported as a result of infection by human papillomavirus (HPV) (eg, HPV 16), and the consequence of the retinoblastoma (Rb) protein inactivation by HPV E7 protein. However, to our knowledge, there have been no studies on the relation between p16(INK4a) overexpression associated with HPV and small cell carcinoma of the cervix, which behaves more aggressively clinically than squamous cell carcinoma. The purpose of this study was to determine whether p16(INK4a) is overexpressed in small cell carcinoma, and if p16(INK4a) is overexpressed, the types of HPV that are related to this cancer. We reviewed 10 cases of small cell carcinoma and examined them for p16(INK4a) overexpression by immunohistochemistry. We also performed HPV typing with polymerase chain reaction (PCR)-sequencing analysis and in situ hybridization and found that p16(INK4a) was overexpressed in every case. PCR-sequencing analyses revealed that all cases were HPV-positive and that 9 cases were positive for HPV 18. Five of the 9 cases positive for HPV 18 were also positive by in situ hybridization and yielded a punctate signal, considered to represent the integrated form. In conclusion, p16(INK4a) was overexpressed and HPV 18 was frequently detected in an integrated form in small cell carcinoma. Therefore, inactivation of Rb protein by HPV 18 E7 protein may be associated with carcinogenesis of small cell carcinoma the same as inactivation of Rb protein by HPV 16 E7 protein is associated with carcinogenesis of squamous cell carcinoma.     

人乳头瘤病毒16型变异株E7蛋白在HeLa细胞中的表达及亚细胞定位

目的构建增强绿色荧光蛋白(EGFP)-人乳头瘤病毒16型变异株E7(HPV16-HBE7)重组质粒pEGFP-HBE7,研究HPV16-HBE7蛋白亚细胞定位,为进一步了解其生物学功能奠定基础。方法采用分子克隆技术,将HPV16-HBE7基因克隆在pEGFP-C1表达载体上,用脂质体法导入宫颈癌细胞中;West-ern印迹检测HBE7蛋白的表达;同时借助免疫荧光技术和EGFP-融合蛋白技术,采用激光共聚焦显微镜观察HBE7蛋白的亚细胞定位。结果重组质粒经PCR、酶切和测序鉴定,其目的片段大小、插入位点和核苷酸序列完全正确;结果表明,转染细胞HBE7蛋白的相对表达量其胞浆明显多于胞核;各个时间段HBE7蛋白均以胞浆分布为主,绿色荧光密集点状分布于细胞浆内,而野生株E7(WE7)蛋白分布在核内。结论人乳头瘤病毒16型变异株E7蛋白主要分布在细胞浆内,以胞浆为主的分布可能和HBE7基因发生突变后丢失核定位信号有关。     

DC—LMP2与rAd—LMP2所诱导的细胞免疫应答效果研究

目的比较DC-LMP2与rAd—LMP2在BALB／c小鼠中的免疫效果。方法从BALB／c小鼠中分离骨髓细胞,在细胞因子作用下体外诱导培养获得小鼠树突状细胞（mDC）,再以100MOI的含EB病潜伏膜蛋白2的重组腺病毒（rAd—LNP2）感染,获得DE—LNP2,免疫酶法确定LMP2在DC中表达。于0,2,4周,分别以PBS、DC-LMP2和rAd-LMP2肌肉免疫BALB／c小鼠,第5和8周检测小鼠体内LMP2特异性细胞应答水平。结果骨髓细胞体外成功诱导获得DC,LMP2在DC中表达。DC—LMP2和rAd—LMP2均可诱导LMP2特异性细胞免疫应答,随时间推移而减弱。rAd—LMP2诱导特异性细胞免疫应答水平高于DC—LMP2。结论肌肉免疫rAd—LMP2能够在BALB／c小鼠中诱导较强的LMP2特异性细胞免疫应答。     

人乳头瘤病毒18型L2表位的定位

人乳头瘤病毒１８例的晚期开放读码框架２编码的蛋白具有型特异性。质粒Ｐ１８Ｌ２００１含有ＰＨＶ１８２ＯＲＦ的全序列，经诱导表达产生的融合蛋白能与ＨＰＶ１８感染的患者血清呈阳性免疫反应。采用Ｈｅｎｉｋｏｆｆ改良的单方向缺失核酸链方法，制备了一组从３＇端至５＇端逐步缩小目的ＤＮＡ分子量的质粒，然后表达一系列从Ｃ端至Ｎ端逐步缩小的融合蛋白。用这一系列融合蛋白与相应血清作Ｗｅｓｔｅｒｎｂｌｏｔ实验，最小的阳     

Human papillomavirus-16 and -18 replication in esophagus squamous cancer cell lines does not require sheterologous E1 and E2 proteins

Human papillomaviruses (HPVs) are doublestranded DNA viruses that replicate in the nuclei of squamous epithelial cells. HPVs can be classified into high-risk (e.g., types 16, 18, 31, and 33) or low-risk (e.g., types 6, 11, and 30), depending on their association with benign or malignant tumors. We recently described the association of HPV-16 and -18 with esophagus squamous cell cancer. HPV replication was studied in representative cell lines derived from esophagus cancers. HPV-16 and -18 genomes were independently transiently transfected into HCE-4 and HCE-7 cell lines with and without E1 and E2 genes under heterologous promoters. Southern blot analysis demonstrated that these cell lines support viral replication. However, heterologous E1 and E2 are not required for HPV replication. These findings suggest that specific host nuclear factors in esophageal squamous epithelial cells may support HPV replication. © 1995 Wiley-Liss, Inc.     

Induced apoptosis and expression of related proteins in granulosa cells from women undergoing IVF: a preliminary study

BACKGROUND: Because apoptosis may be involved in the outcome of IVF, the expression of pro- and anti-apoptosis proteins in a model of induced granulosa cell (GC) apoptosis was evaluated in 25 women with normal FSH levels undergoing IVF. METHODS: After 1 day of culture, apoptosis was induced with interferon (IFN)-gamma (200 IU/ml), followed 24 h later by an agonistic anti-Fas antibody (0.5 microg/ml). On day 3, apoptotic GC, identified by chromatin condensation and/or nuclear fragmentation after DAPI staining, were counted among 1000 cells in randomly chosen fields under UV microscopy, and enabled allocation of women into two groups with either low (group 1) or high (group 2) percentages of apoptosis (11.6 +/- 4.8 and 59.5 +/- 14.8% respectively; P < 0.001). Immunoblotting was used to evaluate the following in proteins: poly (ADP-ribose) polymerase (PARP), caspases 8 and 3, Bcl-2, heat shock protein (HSP) 70, Bax, Bak and Stat-1 (a protein known to be inducible by IFN-gamma). RESULTS: Based on densitometric analysis of immunoblots, the PARP 116 kDa bands were respectively 4.3- and 33.3-fold lower for treated groups 1 and 2. Caspase 8, caspase 3 and HSP70 were expressed slightly less in treated group 2 than treated group 1. Densitometric analysis of bands corresponding to Bcl-2 showed respectively for treated groups 1 and 2, 3.2- and 2.5-fold decreases. Bak expression was similar in both control groups, and comparably lower in the two treated groups. With regard to Stat-1, densitometry showed 3.3- and 1.3-fold increases respectively in treated groups 1 and 2. CONCLUSIONS: These results suggested that Fas-mediated apoptosis of GC is accompanied by significant changes in proteins acting in apoptosis, and that this type of programmed cell death might play a potential prognostic role for women undergoing IVF.