Inducible heat shock protein 70 enhances human papillomavirus type 31 genome replication, viral capsid protein nuclear localization and progeny virion morphogenesis in human keratinocytes

Human papillomaviruses (HPVs) are small, non-enveloped double stranded DNA viruses that demonstrate a strict species and cell type tropism for human epithelial cells. The association between high-risk HPV types and cervical cancer is well established. Additionally, HPVs have been implicated as causes in development of several other epithelial cancer types. Increasing data indicate heat shock proteins (HSPs) including inducible HSP70 (HSP70i) are involved in the replicative cycles of different viruses including adenoviruses, polyomaviruses (PyV), and some RNA viruses. Cell-free system studies implicate HSP70i in HPV11 genome replication with E1 and E2 proteins, and there is evidence that HSP70 is involved in capsid assembly and disassembly for PyV and PV. HSP70 expression is increased in HPV16 E6/E7 gene transduced human primary keratinocytes, and frequently detected in early stage uterine cervical cancer at levels in conjunction with lesion severity. In this study we carry out analyses with the natural host cell to assess HSP70i\u27s role in the viral infectious life cycle. For these studies we used the organotypic (raft) culture system to recapitulate the full viral life cycle of the high-risk human papillomavirus type 31 (HPV31). Upon heat shock of HPV31 infected organotypic tissues, we find high and sustained expression of HSP70i coincident with enhanced HPV genome replication and virion production. Whereas there is no detectable effect on total L1 expression levels, we find that HSP70i interacts with L1, colocalizes with and enhances L1 nuclear localization in differentiated cells. Adenovirus-mediated gene transfer was used to study the effects of HSP70i in naturally HPV infected differentiating tissues and showed results similar to those in heat shocked rafts. In HPV31 infected monolayer cells ectopically expressing viral capsid proteins, without obvious impact on L1 expression levels, our results suggest wild type HSP70i interacts with and promotes L1 translocation into nucleus concomitant with increased HPV genome replication and virion production. Alternatively, HSP70i ATPase domain mutant (HSP70i(K71A)) impedes virion production while viral genome levels, L1 expression and localization demonstrate the same pattern as control. These results indicate that HSP70i is involved in diverse aspects of the viral life cycle including genome replication, capsid protein transportation and virion morphogenesis. We conclude that HSP70i contributes directly to these HPV replicative viral activities and the production of infectious progeny virions

Assessment of Human Papillomavirus in Lung Tumor Tissue

Background Lung cancer kills more than 1 million people worldwide each year. Whereas several human papillomavirus (HPV)-associated cancers have been identified, the role of HPV in lung carcinogenesis remains controversial. Methods We selected 450 lung cancer patients from an Italian population-based case-control study, the Environment and Genetics in Lung Cancer Etiology. These patients were selected from those with an adequate number of unstained tissue sections and included all those who had never smoked and a random sample of the remaining patients. We used real-time polymerase chain reaction (PCR) to test specimens from these patients for HPV DNA, specifically for E6 gene sequences from HPV16 and E7 gene sequences from HPV18. We also tested a subset of 92 specimens from all never-smokers and a random selection of smokers for additional HPV types by a PCR-based test for at least 54 mucosal HPV genotypes. DNA was extracted from ethanol- or formalin-fixed paraffin-embedded tumor tissue under strict PCR clean conditions. The prevalence of HPV in tumor tissue was investigated. Results Specimens from 399 of 450 patients had adequate DNA for analysis. Most patients were current (220 patients or 48.9%) smokers, and 92 patients (20.4%) were women. When HPV16 and HPV18 type-specific primers were used, two specimens were positive for HPV16 at low copy number but were negative on additional type-specific HPV16 testing. Neither these specimens nor the others examined for a broad range of HPV types were positive for any HPV type. Conclusions When DNA contamination was avoided and state-of-the-art highly sensitive HPV DNA detection assays were used, we found no evidence that HPV was associated with lung cancer in a representative Western population. Our results provide the strongest evidence to date to rule out a role for HPV in lung carcinogenesis in Western populations

Portable 3D Gait Analysis Assessment in MTT Treat Chronic Ankle Instability: A Retrospective Study

Purpose. Retrospective analysis of the effect of portable 3D gait analysis as an innovative evaluation method in the treatment with MTT on chronic ankle instability patient. Methods. From January 1, 2019, to December 31, 2019, 56 cases of chronic ankle instability (CAI) were extracted from the medical record system of Shenzhen Longhua District Central Hospital. All the patients of 56 cases accepted the medical training therapy (MTT). As outcome parameters, the alterations of the Cumberland ankle instability tool (CAIT), foot and ankle ability measure (FAAM), were used before the treatment and after treatment; meanwhile, the portable apparatus 3D gait analysis was used to measure the gait parameters. Conclusion. The results showed only ankle angle parameters Y-axis, maximum dorsiflexion during support period (°) had a significant difference, and the p value is 0.039. Meanwhile, the CAIT, FAAM, and most 3D gait analysis data had no significant difference. This particular statistical difference shows that CAI can be measured scientifically and objectively, although most measurement parameters have no change. These results make further reveal that the CAI patients are suffering with dynamic abnormality of ankle motion angle; this also provides us with a measurable and systematic evaluation reference plan for CAI treatment in the future

Modulating the gut microbiota is involved in the effect of low-molecular-weight Glycyrrhiza polysaccharide on immune function

ABSTRACTLow molecular weight (6.5 kDa) Glycyrrhiza polysaccharide (GP) exhibits good immunomodulatory activity, however, the mechanism underlying GP-mediated regulation of immunity and gut microbiota remains unclear. In this study, we aimed to reveal the mechanisms underlying GP-mediated regulation of immunity and gut microbiota using cyclophosphamide (CTX)-induced immunosuppressed and intestinal mucosal injury models. GP reversed CTX-induced intestinal structural damage and increased the number of goblet cells, CD4+, CD8+ T lymphocytes, and mucin content, particularly by maintaining the balance of helper T lymphocyte 1/helper T lymphocyte 2 (Th1/Th2). Moreover, GP alleviated immunosuppression by down-regulating extracellular regulated protein kinases/p38/nuclear factor kappa-Bp50 pathways and increasing short-chain fatty acids level and secretion of cytokines, including interferon-γ, interleukin (IL)-4, IL-2, IL-10, IL-22, and transforming growth factor-β3 and immunoglobulin (Ig) M, IgG and secretory immunoglobulin A. GP treatment increased the total species and diversity of the gut microbiota. Microbiota analysis showed that GP promoted the proliferation of beneficial bacteria, including Muribaculaceae_unclassified, Alistipes, Lachnospiraceae_NK4A136_group, Ligilactobacillus, and Clostridia_vadinBB60_group, and reduced the abundance of Proteobacteria and CTX-derived bacteria (Clostridiales_unclassified, Candidatus_Arthromitus, Firmicutes_unclassified, and Clostridium). The studies of fecal microbiota transplantation and the pseudo-aseptic model conformed that the gut microbiota is crucial in GP-mediated immunity regulation. GP shows great potential as an immune enhancer and a natural medicine for treating intestinal inflammatory diseases