The viral genome editing technologies have been shown to largely contribute to the stud-ies of viral gene functions and to the treatment of infectious diseases. With the development of genetic engi-neering technologies, a variety of viral genome mutation technologies have been established. The homologous recombination is traditionally used to edit the viral genome, but the low frequency of homologous recombina-tion limits its application. Large genome DNA viruses including herpes simplex virus 1 can be edited by bac-terial artificial chromosome system. However, the cloning of viral genome to bacterial artificial chromosome plasmid is both laborious and time-consuming, and parts of the plasmid genes or drug selection markers may remain in the genome of virus and then affect the function of virus. Fortunately, the CRISPR-cas9 system which was discovered in 2013 can be used to edit the viral genome efficiently and accurately. Furthermore, the experiment is simple and will not leave any trail of the viral genome when the genetic mutation is per-formed. Here, we briefly review the application of homologous recombination, bacterial artificial chromosome system and CRISPR-cas9 system in researches on herpes simplex virus 1.

The molecular modifications of Herpes Simplex Virus Type Ⅰ (HSV-1) proteins represented by acetylation and phosphorylation are essential to its biological functions.The cellular chromatin-remodeling/assembly is involved in HSV-1 associated gene transcriptional regulation in human cells harboring HSV-1 lytic or latent infections.Further investigation on these biological events would provide a better understanding of the mechanisms of HSV- 1 viral gene transcriptional regulation.

Herpes simplex virus type 1 (HSV-1), as a pathogen usually leading to herpes labialis, has been found to cause more cases of genital herpes and be associated with the occurrence of neurological diseases such as Alzheimer′s disease (AD). AD is a chronic and progressive degenerative disease of the central nervous system, the pathogenesis of which has not been fully elucidated, and currently there is a lack of drugs that can effectively treat the disease. The hypothesis that HSV-1 infection is involved in the pathogenesis of AD has been discussed for a long time, but there is no definite evidence to prove it. This article briefly reviewed the research progress in the pathogenesis of AD caused by HSV-1.

Nuclear proteins often form punctiform structures, but the precise mechanism for this process is unknown. As a preliminary study, we investigated the aggregation of an HSV-1 immediate-early protein,infected-cell protein 22 (ICP22), in the nucleus by observing the localization of ICP22-EGFP fusion protein.Results showed that, in high-level expression conditions, ICP22-EGFP gradually concentrates in the nucleus,persists throughout the cell cycle without disaggregation even in the cell division phase, and is finally distributed to daughter cells. We subsequently constructed a mammalian cell expression system, which had tetracyclinedependent transcriptional regulators. Consequently, the location of ICP22-EGFP in the nucleus changed with distinct induction conditions. This suggests that the cellular location of ICP22 is also influenced by promoter regulation, in addition to its own structure. Our findings provide new clues for the investigation of transcriptional regulation of viral genes. In addition, the non-protease reporter system we constructed could be utilized to evaluate the role of internal ribosome entry sites (IRES) on transcriptional regulation.

Objective To study the diagnosis and treatment of malignant renal angiomyolipoma with metastases. Methods Three cases of advanced malignant renal angiomyolipoma with metasta-ses were retrospectively reviewed. Case 1 was a 55-year-old woman presenting with recurrent low-grade fever and aching pain in left flank. Ultrasound showed solid mass in the left kidney. Left radical nephrectomy was then performed. The right pulmonary lobectomy of the inferior lobe and wedge ex-section of superior lobe was performed 7 years later because of multiple pulmonary metastases. Case 2 was a 37-year-old woman. Left nephrectomy was performed because Ultrasound and CT showed left kidney solid mass. Six years later, multi-site metastases were found in liver and retroperitoneum and mestastasis tumors were resected. At 10 years after the primary diagnosis, CT showed multi-metasta-ses in liver and retroperitoneum. The retroperitoneal masses were resected and liver lesions were trea-ted by radiofrequency ablation. Case 3 was a 34-year-old man presenting with swelling pain in right flank. CT scan showed a lesion in the right kidney and right radical nephrectomy was performed. Four months after the surgery, MRI revealed multiple liver and retroperitoneal nodules. All the 3 cases had not been diagnosed with tuberous sclerosis and did not accept chemotherapy. Results The cut sur-face of the lesions was red-brown and yellow and the texture was tender. Under microscopic examina-tion, the tumors of case 1 and case 3 were composed of sheets or nests of large polygonal epithelioid cells. It revealed that occasionally clear cytoplasm with abundant eosinophilic, prominent nucleoli, and multinucleated and markedly pleomorphic form. Necrosis was presented as well. Large areas of case 2 tumor were made up of spindle smooth-muscle cells, adipose tissue, thick-wall blood vessels and some areas merged with a proliferation of epithelioid which was consistent with typical angiomyolipoma. Im-munohistochemical study showed that the epithelioid cells and spindle smooth-muscle cells were posi-tive for VM, HMB45, Melan-A and negative for S100, CK. Case 1 and case 3 were diagnosed with malignant epithelioid angiomyolipoma, while case 2 was diagnosed with malignant classic angiomyoli-poma and epithelioid in part of the tumor. Case 1 was well alive. Case 2 was alive with tumor 12 years after the diagnosis. And case 3 was missed in the follow-up 3 months after metastasis resection. Conclusions Malignant renal angiomyolipoma is a rare disease. The diagnosis depends on histopatho-logic, immunohistochemieal study and clinical follow-up. Radical resection of the primary, recurrent and metastatic tumors is the main therapy. It needs more research to clarify if metastasis has any effect on prognosis.

The three immediate-early proteins of HSV-1, ICPO, ICP22, and ICP27, have specific and pivotal functions in transcriptional activation and inhibition, multiple regulatory and control processes of viral genes. In this paper, the expression and localization of these three proteins were studied in neuroblastoma cells using biochemical assays, and their possible and potential interactive functions are discussed. The data show that the three proteins are localized in different structures, specifically in the PML-NB-associated structure, which is a specific nuclear structure composed of many protein molecules and bound tightly to the nuclear matrix in neuroblastoma cells. The results suggest that the activating and suppressive functions of ICPs are mostly dependent on their transcriptional and regulatory roles, including the PML-NB-associated structure.

Purpose　The aim is to invest the mechanism and activity of kringle1～3 gene of angiostatin. Methods　Kringle1～3 gene of angiostatin was abtained from hepatocyte of normal human by RT-PCR and inserted into expression vector pPIC9K in Pichia expression system. The recombinant protein was induced secrete , purified and test the activity by MTT.Results　　Krigle1～3 protein could inhibit vascular endothelial cell proliferative activity.Conclusion　The reaction similar apoptosis induced by kringle1～3 recombinant protein of angiostatin inhibit cell proliferation.

Objective To express and purify the recombinant UL7 protein of herpes simplex virus 1 (HSV-1), to prepare the corresponding UL7-specific polyclonal antibody and to preliminarily analyze the expression of UL7 protein during the proliferation of HSV-1. Methods The UL7 gene was amplified by PCR and then cloned into the pGEX-5X-1 vector for expression of UL7 protein in the prokaryotic expression system. The constructed expression plasmid, pGEX-5X-1-UL7, was transformed into E. coli BL21 (DE3) to induce the expression of UL7 protein by IPTG. The purified GST-UL7 fusion protein was used as antigen to inject the ICR mouse for the preparation of polyclonal antibody specific for UL7 protein. The titer and speci-ficity of the polyclonal antibody were analyzed by using indirect ELISA and Western blot assay, respectively. The UL7 protein-specific polyclonal antibody was used to detect the expression of UL7 protein at different time points after infecting Vero cells with HSV-1. Results The GST-UL7 fusion protein was efficiently ex-pressed in E. coli BL21 (DE3). The UL7 protein-specific polyclonal antibody was prepared with high titer (1 ∶ 105) and high specificity as indicated by the indirect ELISA and Western blot assay. The expression of UL7 protein was detected at different time points after infecting Vero cells with HSV-1. Conclusion The GST-UL7 fusion protein was obtained successfully and the UL7 protein-specific polyclonal antibody was pre-pared. Accompany with the proliferation of HSV-1, the expression of UL7 protein was detected at different time points by using the polyclonal antibody.

Enterovirus 71 (EV71) is a common cause of Hand, foot, and mouth disease (HFMD) and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences (891 nucleotides) from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous to members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine(H) at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains.

The complete nucleotide sequence of the mumps virus SP, which was isolated in China, was determined. As with other mumps viruses, its genome was 15 384 nucleotides (nts) in length and encoded seven proteins. The full-length nucleotide sequence of the SP isolate differed from other strains by 4%-6.8% at the nucleotide sequence level. Due to variations of amino acids over the full genome (including the HN and N genes), this isolate exhibited significant variations in the antigenic sites. This report is the first to describe the full-length genome of a genotype F strain and provide an overview of the diversity of genetic characteristics of a circulating mumps virus.